LOCUS       sequence-48050-e        9539 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 9539)
  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 9539)
  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: pCFJ212 - pDESTR4-R3(cxTi10816, IV)
FEATURES             Location/Qualifiers
     source          1..9539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(151..170)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(835..1344)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1030..1051)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1238..1257
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      1455..2043
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1944..1963
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    2104..2125
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2140..2145,2146..2163,2164..2170)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2178..2194
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2183..2205
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2202..2218
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2202..2218
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(3808..3819)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    5885..6008
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(6049..6354)
                     /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(6151..6170)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(6696..7355)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     7270..7289
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(7409..7415,7416..7433,7434..7439))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(7464..7587)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(7595..7611)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9130..9146)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9139..9161)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        9367..9471
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9472..9539,1,2..793)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
       61 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
      121 ttctccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga
      181 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
      241 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
      301 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
      361 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
      421 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
      481 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
      541 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
      601 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
      661 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
      721 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
      781 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaccccg
      841 gttgataatc agaaaagccc caaaaacagg aagattgtat aagcaaatat ttaaattgta
      901 aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
      961 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagcccga gatagggttg
     1021 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     1081 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc caaatcaagt
     1141 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     1201 agagcttgac ggggaaagcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     1261 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     1321 ttaatgcgcc gctacagggc gcgtaaaagg atctaggtga agatcctttt tgataatctc
     1381 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     1441 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     1501 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     1561 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     1621 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     1681 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     1741 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     1801 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     1861 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     1921 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     1981 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     2041 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     2101 atgtaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     2161 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     2221 tgggtcaccc aaggcgacaa cgtaagtgac tgaaggcatt aaaggtacat gcgtccgcgc
     2281 aagtgtccgc ttttgctggc ctaacaagta agagaaacaa gtaagaaaga gtaagagaaa
     2341 actcacccga ggtgtctctt ctagcaaccg cctattcaga taattttaag ccagttcgag
     2401 agttatttgt cgcatggtat agaagctcgt tatttacatt aaatttttag tcttcccgca
     2461 gaagtggacg atgcgatggg ctacttcaaa aatctgacgc cacgtgtcaa ggaccttcac
     2521 aagagcatga caaacttgga agatattagc aagtggcaga agaaggccag tttctctggc
     2581 acccttgaga attactcgcg tctcggtgac aagatcaatg tgaaaccatt tatggatgct
     2641 gttgatgcta gaatgggtgc cgaagctgat gccgtgaaag gggtataaag ctactgttgc
     2701 acgacgactc acagactaat cattccaggt cctcgcgatt tgtgaaaaat acaagtcatt
     2761 ctaccaaaac gagggaaaac ttcacgcgga cagtatcgcc aatttgaata ggactcggct
     2821 cgacatggac agtgcggcgg ataaatatgc gaacaacgag actgaagtta acaagactcg
     2881 tttggataac agtaccacgg aatttgaagt ggcttgtgag aggtttgtag gtctttcgta
     2941 ggcacgatat ccttatttga atgtgtttca gaatgcgaga actggcgaac ggaatcaaga
     3001 caattgaatc gaaccattct tcctggcaag acggtcttat gaaggagatt aaagtggcgt
     3061 tgcgtaaata aagaaaggat cctctttgaa aataaagatt ttctacattc acttattttt
     3121 ttagctttat acttttcaga tcgaatgaac tgtttccaac gtttccatat taattttttt
     3181 tagttaaaaa agttgggtat taacagtttt gttaacagtt ttgtcttccg acagatgtga
     3241 gacggaaaaa tggccaaaat tcgagatttt gactaaaatc agtgtagctt tttcaaaatt
     3301 ttgaaccggc atgaaaaagt ttgaaaagtt ttcgagaagt ttcaatccgt tcatttgagc
     3361 ccatgttggg tctataagat caaaatcgcc cgaactgtta gtcctccttt aagttacgat
     3421 tattaacaaa atgtcgccta agtttagttt cacacttgtc tcacactagt cgatgaactg
     3481 tgtgactact gtagtgagta cagtagggcc cgtgtaaaaa tacaaaaacc aagaaaaagt
     3541 aaaccaaaat aaaaaacgaa tttattgttc actacaacca aactgcaaag ttcttttctt
     3601 acccatcagc tcaatcgtgt acttgcgtgg aaacaaatat ggtagcaaac tcacttcgtc
     3661 tttgagccaa tttatccaag tccttgtaaa aagtattcga aaattgttaa cggataaaat
     3721 gtttattata atatcaaaaa caatttgtca gttgaccact ttttgatata tttgacagaa
     3781 accgggatga attggctcaa agttagggcg cccttctatt acagggtttc tgataacaaa
     3841 ccggttatta actcccaaca agggatgatt tcaattcatc atgctcaaat tgacccaaat
     3901 taagttacat gacaaattca tcgccctttt caactcttct tggctcatca tctgttattc
     3961 tgttcctatt ctgtctgcac cctatacccc tttgcatact ctctcgtcat tcctttcttg
     4021 gatagtgtct tcttctcccc agctctgcta cttctatgac ttgcgcccgc tgcttttccg
     4081 ctcgttcctc tcctctctga cgtcttcttc tgcttctttt ctccttcctt tttcccttct
     4141 caatcttcct ctttttccca tttctgtcaa tcattcgaag aagaagaaga agaagaccct
     4201 cattcatttc attttttttt ctgtcggtgt gtgctgctcg gttaagagtg agctctctat
     4261 tccacgtctt cttctttttc ttcttcgatt cgaatcaatc actccaccaa aacgcattcg
     4321 tttttgggat tcaccccgcg gttcgcaata ggtttctttt tcaaattatt agcgttataa
     4381 atagaaaaat ggtggagttt caataaaaat gataatttac aaagtgtatt ttgattacat
     4441 gtactcaaaa aggttgaaat ttttcatacc agttttccgg aaatccatct gatatcatta
     4501 tcgtattttt ctttttaaaa atgttttcaa aaaaaaacaa aatatagctg gttatttggc
     4561 accctctaat taccattttc ctgtcacacc acactctttt ccttcttcca cttctttccg
     4621 ttttcagccg cttccaacca aaccgatatg aaagccgagc aacaacgatc gattccaccc
     4681 ggctcggcga ccttcccgtc gcaggtgaga ctcagaaaac tagagaaacc gctcaactaa
     4741 ctcttgatat ccgatttcat tcttttctct tttctttttg ttgaactttt ccatatttcc
     4801 agatgccacg gccaccacca agcaccgaac aaggaatcac aacggaatcg gagcttgcga
     4861 agaaagctca aatcactccg aacgacgttt tagcacttcc gggaatcact caaggtatct
     4921 tgctcccttt ggatttcagt ttaaaacata aatttaggat tcttatgctc cccatctgcg
     4981 aacatctata acatcgagtt caccaagttc caaatccgtg atctggacac tgagcaagtg
     5041 cttttcgaga tcgccaaacc ggagaacgat caggagaatg atgagtcgcc acaggagtcg
     5101 gcaagatacg tgcgttatag atttgctcca aactttttga aactcaaaac ggtcggagca
     5161 actgtggaat tcaaagtagg agacatccca atccatcatt tccgaatgat cgaacgtcac
     5221 ttcttcaaag atcgccttct gaagtgtttt gactttgaat tcggattctg tattccgaat
     5281 tcacgaaaca actgtgaaca tatctatgag ttccctcaac tctctcaaca actcagtgag
     5341 tcattattct aaaaagtaca attcaaaaga ctaatctctt tttcagtgga cgacatgatc
     5401 aacaatccaa acgagactcg ttctgacagc ttctatttcg tcgataacaa actcgtcatg
     5461 cacaacaaag ccgactactc atatgatgca taaatattta atacaaaaat gttctggata
     5521 attattctgt cgaatagaaa aaaaactcca aatgtgatta aattccaata attcctgtct
     5581 agtttgttcc ttccccttcc ccttctcatg ttcaatgcat tcctaagctt ttcagttccc
     5641 cccttgtttc tatatttttt cgctgtcctg tcacactcgc taaaaacact aatcacacgg
     5701 aaatctgttt tcaataaaaa ctccaacttt aactcatttt caatttcaac tgaaagattt
     5761 tttcattaga gaatgtctag aactaggccc gggctacgta atacgactca cttaaggcct
     5821 tgactagagg gtaccagagc tcacctaggc aggaacagct atgaccatga ttacgccaag
     5881 ctatcaactt tgtatagaaa agttgaacga gaaacgtaaa atgatataaa tatcaatata
     5941 ttaaattaga ttttgcataa aaaacagact acataatact gtaaaacaca acatatccag
     6001 tcactatggt cgacctgcag actggctgtg tataagggag cctgacattt atattcccca
     6061 gaacatcagg ttaatggcgt ttttgatgtc attttcgcgg tggctgagat cagccacttc
     6121 ttccccgata acggagaccg gcacactggc catatcggtg gtcatcatgc gccagctttc
     6181 atccccgata tgcaccaccg ggtaaagttc acgggggact ttatctgaca gcagacgtgc
     6241 actggccagg gggatcacca tccgtcgccc gggcgtgtca ataatatcac tctgtacatc
     6301 cacaaacaga cgataacggc tctctctttt ataggtgtaa accttaaact gcatttcacc
     6361 agcccctgtt ctcgtcggca aaagagccgt tcatttcaat aaaccgggcg acctcagcca
     6421 tcccttcctg attttccgct ttccagcgtt cggcacgcag acgacgggct tcattctgca
     6481 tggttgtgct taccgaaccg gagatattga catcatatat gccttgagca actgatagct
     6541 gtcgctgtca actgtcactg taatacgctg cttcatagca tacctctttt tgacatactt
     6601 cgggtataca tatcagtata tattcttata ccgcaaaaat cagcgcgcaa atacgcatac
     6661 tgttatctgg cttttagtaa gccggatcct ctagattacg ccccgccctg ccactcatcg
     6721 cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga
     6781 tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg
     6841 gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa
     6901 ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag
     6961 gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa
     7021 tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg
     7081 taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc catacggaat
     7141 tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc
     7201 ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag
     7261 gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata
     7321 tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat
     7381 ctcgacggat cctaactcaa aatccacaca ttatacgagc cggaagcata aagtgtaaag
     7441 cctggggtgc ctaatgcggc cgccatagtg actggatatg ttgtgtttta cagtattatg
     7501 tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt
     7561 ctcgttcaac tttattatac atagttgata attcactggc cgtcgtttta cactcgagac
     7621 gtacggtgcg cgcgatgcat tcgaagatct gcccactagt gagtcgtatt ataagtgcaa
     7681 gtaagatcag tgtttgtatt taagacacac ggccaaattt cttattttcc tgctattttc
     7741 tccaaatatg attaaaattt gttcatttca aaattgcctc caacttgttt tggggcaaaa
     7801 atttcacaaa acttcacagc aaaatgtgaa acaatattgc tatctattga aaacattaat
     7861 gatagtcgag tgacttcttg aaccaatact catcgcatca aatgcctttt ctcgaatact
     7921 ctgtaataat tggcatataa aaacatacat actcatcagt ttgaaagcta acctttacca
     7981 taatcattac taaaattgga tttaaagccg gacaccaaga aaaaagatac ggtagataaa
     8041 ctgacagatg agaagtctcg atacgaaata atggcattag aaaagacaca tagaatggaa
     8101 tgtggatagt gagaaatggt agaatgaact gaaaaattga gaatttaagg agaaatgtta
     8161 tttttaagct gctttgcttg aaactttgaa tttaaaattt tagaacaatt actgtttcaa
     8221 aatattttta tttttattta ctccatttga aaaaaaagca acattgcttg ttcaatttct
     8281 gttattgcct tacctgaacc ataagtacca tattcaattg ttgttgagac ttcatcgaat
     8341 ttgtactccg acttccctta tgaatttcgt atctgataat tgtccaacaa accatcgcaa
     8401 ttactgtagt tatcactaga acagccatta tataaatcgt acatgcccat gaaattgggt
     8461 cagcaacaaa gtcattcaac gtgtaatcat atctttctag tgatgcaatt aaatatgcag
     8521 attttccgat tccattcaga ttgtatacga tatattctga tagttgatcg tcagcagcat
     8581 ctgggacata gccaaaggtg attagaactg tggatattag agagacaata aatgatatga
     8641 ttgtggttgt aaggaccacg tttttttgtg ttctcgttgt tctgaaagta taatttgtag
     8701 attcttaaac tcagaattta tcatatttga actccgctgt ttttgaatta agtaatagct
     8761 gaaacagtga acattagttt aaaaatataa ttttacaggc acgcacaaac ttcggtgtgg
     8821 gtacgtatta cctgcctcct attgaatatt cattgtttaa acgttttcag aaaaaaagtg
     8881 tacatgtttt ttggagtatg atgatgatgt agtcgcaagg cagatatgaa atgtaggtaa
     8941 cacagtgcac agatgcgttt tttgccaaca tcaagggttg taagtaactt ttcagagaaa
     9001 aacatataga aattttgttt caagtatcgt atcagttttt ttctgattaa cttataaaaa
     9061 gaactcttcg acaatttgaa tgcttaattt aaaaaaatta agtcgaatat ttcagcatgc
     9121 caatgatcaa ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     9181 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     9241 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgct tcgcttggta
     9301 ataaagcccg cttcggcggg cttttttttg ttaactacgt caggtggcac ttttcgggga
     9361 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     9421 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     9481 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgc
//
