LOCUS       sequence-48054-e        8130 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 8130)
  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 8130)
  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: pCJF604 - pDESTR4-R3(ttTi4391, I)
FEATURES             Location/Qualifiers
     source          1..8130
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      155..743
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     644..663
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1279..1295
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1279..1295
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    1315..1438
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1479..1784)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(1581..1600)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(2126..2785)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2700..2719
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(2839..2845,2846..2863,2864..2869))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(2894..3017)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3025..3042)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3025..3041)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3191..3202)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6884..6903)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7003..7025
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7063..7081)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7149..7253
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7254..7322,7323..8114)
                     /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(7472..7491)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
       61 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
      121 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
      181 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
      241 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
      301 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
      361 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
      421 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
      481 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
      541 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
      601 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
      661 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
      721 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggtct
      781 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
      841 gtattaccgc ctttgagtga gctgatacct gatcaccgtt ttcaacaaga ttacagacat
      901 cgcagtgaaa tcgaggagga gtgtatcaag cagctattga tgattcctcc gatgtatgcg
      961 gattttgact attattattg agaataaagc ttgttataac tcctacagta accccacagt
     1021 acctcgacag tatctctaca gttcctctac aataatctta cagtacccct acagtacctc
     1081 acaataccac ttaggtacct ctacagtacc tctacagtat ccttacagtc cccctacagc
     1141 acccctacag tacttctaca gtactcccat ggtatcctta cagtacccct aaacaataat
     1201 cttgcagtac cgctacagtg ccccttcggt ttctctacgg tatagatgca acacccctac
     1261 tcagtaaaac cgttcataca ggaaacagct atgaccatga ttacgccaag ctatcaactt
     1321 tgtatagaaa agttgaacga gaaacgtaaa atgatataaa tatcaatata ttaaattaga
     1381 ttttgcataa aaaacagact acataatact gtaaaacaca acatatccag tcactatggt
     1441 cgacctgcag actggctgtg tataagggag cctgacattt atattcccca gaacatcagg
     1501 ttaatggcgt ttttgatgtc attttcgcgg tggctgagat cagccacttc ttccccgata
     1561 acggagaccg gcacactggc catatcggtg gtcatcatgc gccagctttc atccccgata
     1621 tgcaccaccg ggtaaagttc acgggggact ttatctgaca gcagacgtgc actggccagg
     1681 gggatcacca tccgtcgccc gggcgtgtca ataatatcac tctgtacatc cacaaacaga
     1741 cgataacggc tctctctttt ataggtgtaa accttaaact gcatttcacc agcccctgtt
     1801 ctcgtcggca aaagagccgt tcatttcaat aaaccgggcg acctcagcca tcccttcctg
     1861 attttccgct ttccagcgtt cggcacgcag acgacgggct tcattctgca tggttgtgct
     1921 taccgaaccg gagatattga catcatatat gccttgagca actgatagct gtcgctgtca
     1981 actgtcactg taatacgctg cttcatagca tacctctttt tgacatactt cgggtataca
     2041 tatcagtata tattcttata ccgcaaaaat cagcgcgcaa atacgcatac tgttatctgg
     2101 cttttagtaa gccggatcct ctagattacg ccccgccctg ccactcatcg cagtactgtt
     2161 gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa
     2221 tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg
     2281 gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg
     2341 gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt
     2401 caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt
     2461 attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt
     2521 gaacactatc ccatatcacc agctcaccgt ctttcattgc catacggaat tccggatgag
     2581 cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct
     2641 ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacattgag
     2701 caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg
     2761 tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat ctcgacggat
     2821 cctaactcaa aatccacaca ttatacgagc cggaagcata aagtgtaaag cctggggtgc
     2881 ctaatgcggc cgccatagtg actggatatg ttgtgtttta cagtattatg tagtctgttt
     2941 tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt ctcgttcaac
     3001 tttattatac atagttgata attcactggc cgtcgtttta cactttgagc caatttatcc
     3061 aagtccttgt aaaaagtatt cgaaaattgt taacggataa aatgtttatt ataatatcaa
     3121 aaacaatttg tcagttgacc actttttgat atatttgaca gaaaccggga tgaattggct
     3181 caaagttagg gcgcccttct attacagggt ttctgataac aaaccggtta ttaactccca
     3241 acaagggatg atttcaattc atcatgctca aattgaccca aattaagtta catgacaaat
     3301 tcatcgccct tttcaactct tcttggctca tcatctgtta ttctgttcct attctgtctg
     3361 caccctatac ccctttgcat actctctcgt cattcctttc ttggatagtg tcttcttctc
     3421 cccagctctg ctacttctat gacttgcgcc cgctgctttt ccgctcgttc ctctcctctc
     3481 tgacgtcttc ttctgcttct tttctccttc ctttttccct tctcaatctt cctctttttc
     3541 ccatttctgt caatcattcg aagaagaaga agaagaagac cctcattcat ttcatttttt
     3601 tttctgtcgg tgtgtgctgc tcggttaaga gtgagctctc tattccacgt cttcttcttt
     3661 ttcttcttcg attcgaatca atcactccac caaaacgcat tcgtttttgg gattcacccc
     3721 gcggttcgca acaggtttct ttttcaaatt attagcgtta taaatagaaa aatggtggag
     3781 tttcaataaa aatgataatt tacaaagtgt attttgatta catgtactca aaaaggttga
     3841 aatttttcat accagttttc cggaaatcca tctgatatca ttatcgtatt tttcttttta
     3901 aaaatgtttt caaaaaaaaa caaaatatag ctggttattt ggcaccctct aattaccatt
     3961 ttcctgtcac accacactct tttccttctt ccacttcttt ccgttttcag ccgcttccaa
     4021 ccaaaccgat atgaaagccg agcaacaaca atcgattcca cccggctcgg cgaccttccc
     4081 gtcgcaggtg agactcagaa aactagagaa accgctcaac taactcttga tatccgattt
     4141 cattcttttc tcttttcttt ttgttgaact tttccatatt tccagatgcc acggccacca
     4201 ccaagcaccg aacaaggaat cacaacggaa tcggagcttg cgaagaaagc tcaaatcact
     4261 ccgaacgacg ttttagcact tccgggaatc actcaaggta tcttgctccc tttggatttc
     4321 agtttaaaac ataaatttag gattcttatg ctccccatct gcgaacatct ataacatcga
     4381 gttcaccaag ttccaaatcc gtgatctgga cactgagcaa gtgcttttcg agatcgccaa
     4441 accggagaac gatcaggaga atgatgagtc gccacaggag tcggcaagat acgtgcgtta
     4501 tagatttgct ccaaactttt tgaaactcaa aacggtcgga gcaactgtgg aattcaaagt
     4561 aggagacatc ccaatccatc atttccgaat gatcgaacgt cacttcttca aagatcgcct
     4621 tctgaagtgt tttgactttg aattcggatt ctgtattccg aattcacgaa acaactgtga
     4681 acatatctat gagttccctc aactctctca acaactcagt gagtcattat tctaaaaagt
     4741 acaattcaaa agactaatct ctttttcagt ggacgacatg atcaacaatc caaacgagac
     4801 tcgttctgac agcttctatt tcgtcgataa caaactcgtc atgcacaaca aagccgacta
     4861 ctcatatgat gcataaatat ttaatacaaa aatgttctgg ataattattc tgtcgaatag
     4921 aaaaaaaact ccaaatgtga ttaaattcca ataattcctg tctagtttgt tccttcccct
     4981 tccccttctc atgttcaatg cattcctaag cttttcagtt ccccccttgt ttctatattt
     5041 tttcgctgtc ctgtcacact cgctaaaaac actaatcaca cggaaatctg ttttcaataa
     5101 aaactccaac tttaactcat tttcaatttc aactgaaaga ttttttcatt agagaatgtc
     5161 tagaactagg cccgggctac gtaatacgac tcacttaagg ccttgactag aggacccaga
     5221 ttcaaaccca tttaaaaccg tacaagccta tactatgaaa aattctaatt ttgggcagat
     5281 gtaaaccaag attgtctaga tccaaactta atttgcatag attgcaagtt tgagtagatg
     5341 taaactgaat ttgtgtaggt ttgtgtagat ccaaattgat tttgtgtaca tccaaattaa
     5401 atttgtgcag ataaattcac aatataggct tgtacggttc taattttgtt tgaatctggg
     5461 tatattctga acgggtttac tgagtacaat gcccccacag tacccctaca gttcttctac
     5521 ataactccta cagtaccctg cttgtttcca agaaactaat tttacaaacc aaaaatattt
     5581 attgcgaaaa atttgaaaat tatcagaaat gttaggcatg tccagcagca gctgcctcct
     5641 tcttcttgcc ctgaaattca attttttttt aaattgtttt tcttcataga aaaagctcac
     5701 cccggctcct ttcttgtgat tttttgctcc tttttttgcg aaaaggtagt tatttggaag
     5761 cgacacgtcc ggggtttcgc cgtgaggccg gcgcggcgct cgacctcata ggatctctaa
     5821 tttgaagagt ttctaattct acgacgctgc aagccttcag acctgcggaa gaacacatta
     5881 gaagtgaaat ccatatctga aaaaagatcg gacaattcaa ccttaaatct aaaacttaca
     5941 agttgtaccc agcccaacaa ttgtccattt tctgccaatc actctggata aagggcttcc
     6001 aaactcatat tccccatgaa agctttcagt aatgactagt ttctcttgta agtctattac
     6061 ctgcaaatat gtacttgctc tttgaaatct attagattca agattaccca aagaagtcga
     6121 tgcttcaatc caacgccgtt atttttccca ttaccattgt cttcataacc atgcacataa
     6181 accccattac caatattaac ttcatctgat agacaaggtg tactgttcgt gaaagtgtca
     6241 tcaaactcga taatcattgg atgataggcc acatgctctg tcatgttcgg gcagtagttc
     6301 aaaatataag ctttcacagg agtagctata tcatggcagt cggtcccgtt cattacacat
     6361 tgaaaacgtc taaacatgtt tttttccaga aatcctgctg gaacctcgac gtatctgaac
     6421 acagattcaa atcaatatta tataacaaat ttagccttac ttcaagtcac ctttacagtc
     6481 taccgattga ttattagccc tccattttct cttacttgac atgacaactc gggaagtagt
     6541 aagaacatgt cttttagata tggcacttgc tccggatata tgataccaga ttccattttc
     6601 atactgtcga gctgtcaagt agaaccattg accattctta ctaagtgcta cattcgcagg
     6661 ttttacattc gcacttgcat tggagtctga tggaccctgg cgttacccaa cttaatcgcc
     6721 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     6781 cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta
     6841 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg
     6901 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     6961 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     7021 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     7081 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     7141 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     7201 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     7261 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     7321 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     7381 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     7441 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     7501 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     7561 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     7621 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     7681 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     7741 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     7801 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     7861 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     7921 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     7981 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     8041 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     8101 ttaagcattg gtaactgtca gaccaagttt
//
