LOCUS       sequence-8986-ex        9131 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 9131)
  AUTHORS   Christian Frøkjær-Jensen, M Wayne Davis, Christopher E Hopkins, 
  TITLE     Single-copy insertion of transgenes in Caenorhabditis elegans
  PUBMED    18953339
REFERENCE   2  (bases 1 to 9131)
  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"
COMMENT     Sequence Label: pCFJ150 - pDESTttTi5605[R4-R3]
FEATURES             Location/Qualifiers
     source          1..9131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(146..734)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(226..245)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(905..1696,1697..1765))
                     /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     1528..1547
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1766..1870)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1938..1956
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1994..2016)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2116..2135
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3836..3853
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3837..3853
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3861..3984
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(4010..4015,4016..4033,4034..4040)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             4094..4753
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4160..4179)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             5095..5400
                     /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     5279..5298
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(5441..5564)
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             7630..7641
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 tgtgaaattg ttatccgctg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
       61 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag accaggaacc
      121 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
      181 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
      241 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
      301 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
      361 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
      421 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
      481 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
      541 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
      601 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
      661 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
      721 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
      781 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
      841 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
      901 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
      961 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     1021 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     1081 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     1141 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     1201 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     1261 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     1321 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     1381 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     1441 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     1501 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     1561 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     1621 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     1681 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     1741 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     1801 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     1861 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     1921 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     1981 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     2041 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     2101 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     2161 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc
     2221 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     2281 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     2341 gtcacgtgtg tttttgaaca cggcgatatg tatctgtaga tcatccccaa ttttcaacaa
     2401 tgaatctaaa tactgaactt tttgtgtaat ctttagtgca gaaaaatgac caaaatcagc
     2461 gattttccga ttatacccaa aatatctcag ctcaatgagc acctataaaa atgtaaaaat
     2521 aaagaaagta gtcgaaccat attttgtaaa ggctaaaaaa ctgaataaat ttttatccat
     2581 ccagcgcggc gttacaatcc gtggcccatg tcacgcacgg ctttcaaaga aaaaaaaaca
     2641 gtctgtttgc gcgacatcga tctggcaaaa atcctaaaaa cgactatgcg tggctcactt
     2701 tatattaata tgaaaaatac aaacttggtg gtgcaggtgc tgcagaggcc atcgatgaag
     2761 aagtggaggc cttgttgtcg atctacaaaa ttgtatggtt gtttgagatt ttccttgaaa
     2821 cgctctcaat cttggatatg atataaattc tggataagat atgatggata tgatctatgt
     2881 ttgatagagt gcatttgaaa ggataacggc cctgcagaca tttttgaaaa aataacaaac
     2941 atttaatttt ctcactcgtt taggctattc ccctattttt gatattcctt cgcacatatg
     3001 aaaactactt tttttcgaaa ctgttaactc cagaatttat aaatctatag cccttacttg
     3061 attatttatt atcatggtta ctcacctcat gtctctttct cttttcaaga agtcttcgtt
     3121 catatctgtt gtctctttcc tccttttgaa tctctgcttt gctcttcgcc attgtttcct
     3181 gaaaataatg taacttgaat tgtgtaatat acttttttaa tttgaatttg gcttgtaacg
     3241 cggaatcact acgtgcggga tcatttctta ctagaaaacc cagaaaatgc catatttcac
     3301 ttatctcggg gtcatttcta attagaaaag ctacaaaaac catttctaat tacggtgact
     3361 ttagaactgg tcgaaatagc taattagaaa tgggtttttg ctaattagag gtgactctat
     3421 gttatctgag taagcctcgt ttgtatgtaa actcacagta ctaataaatg caagacaccc
     3481 gggtttgtct agatatgaaa taattgaaat atcaattctg acagacaata atggtaatct
     3541 tgataaggag ttccacgccc aggagaacac gttagttttc ttgtttttga ttgcgtgcgt
     3601 tattttggag aaaaactcga ttttttacaa aataattttt tgaaaggaac actgttcaat
     3661 aagttttgtc ttttttctca gttgtgatac ggttttttat tcttttttgt agttatacag
     3721 aagaccgtta cgaaacagac tgatatcgaa acaaacactg atcttacttg cacttataat
     3781 acgactcact agtgggcaga tcttcgaatg catcgcgcgc accgtacgtc tcgagtgtaa
     3841 aacgacggcc agtgaattat caactatgta taataaagtt gaacgagaaa cgtaaaatga
     3901 tataaatatc aatatattaa attagatttt gcataaaaaa cagactacat aatactgtaa
     3961 aacacaacat atccagtcac tatggcggcc gcattaggca cccccaggct ttacacttta
     4021 tgcttccggc tcgtataatg tgtggatttt gagttaggat ccgtcgagat tttcaggagc
     4081 taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg
     4141 gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac
     4201 cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc acaagtttta
     4261 tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat tccgtatggc
     4321 aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca ccgttttcca
     4381 tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt tccggcagtt
     4441 tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa
     4501 agggtttatt gagaatatgt ttttcgtctc agccaatccc tgggtgagtt tcaccagttt
     4561 tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca tgggcaaata
     4621 ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtttg
     4681 tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg atgagtggca
     4741 gggcggggcg taatctagag gatccggctt actaaaagcc agataacagt atgcgtattt
     4801 gcgcgctgat ttttgcggta taagaatata tactgatatg tatacccgaa gtatgtcaaa
     4861 aagaggtatg ctatgaagca gcgtattaca gtgacagttg acagcgacag ctatcagttg
     4921 ctcaaggcat atatgatgtc aatatctccg gttcggtaag cacaaccatg cagaatgaag
     4981 cccgtcgtct gcgtgccgaa cgctggaaag cggaaaatca ggaagggatg gctgaggtcg
     5041 cccggtttat tgaaatgaac ggctcttttg ccgacgagaa caggggctgg tgaaatgcag
     5101 tttaaggttt acacctataa aagagagagc cgttatcgtc tgtttgtgga tgtacagagt
     5161 gatattattg acacgcccgg gcgacggatg gtgatccccc tggccagtgc acgtctgctg
     5221 tcagataaag tcccccgtga actttacccg gtggtgcata tcggggatga aagctggcgc
     5281 atgatgacca ccgatatggc cagtgtgccg gtctccgtta tcggggaaga agtggctgat
     5341 ctcagccacc gcgaaaatga catcaaaaac gccattaacc tgatgttctg gggaatataa
     5401 atgtcaggct cccttataca cagccagtct gcaggtcgac catagtgact ggatatgttg
     5461 tgttttacag tattatgtag tctgtttttt atgcaaaatc taatttaata tattgatatt
     5521 tatatcattt tacgtttctc gttcaacttt tctatacaaa gttgatagct tggcgtaatc
     5581 atggtcatag ctgttcctgc ctaggtgagc tctggtaccc tctagtcaag gccttaagtg
     5641 agtcgtatta cgtagcccgg gcctagttct agacattctc taatgaaaaa atctttcagt
     5701 tgaaattgaa aatgagttaa agttggagtt tttattgaaa acagatttcc gtgtgattag
     5761 tgtttttagc gagtgtgaca ggacagcgaa aaaatataga aacaaggggg gaactgaaaa
     5821 gcttaggaat gcattgaaca tgagaagggg aaggggaagg aacaaactag acaggaatta
     5881 ttggaattta atcacatttg gagttttttt tctattcgac agaataatta tccagaacat
     5941 ttttgtatta aatatttatg catcatatga gtagtcggct ttgttgtgca tgacgagttt
     6001 gttatcgacg aaatagaagc tgtcagaacg agtctcgttt ggattgttga tcatgtcgtc
     6061 cactgaaaaa gagattagtc ttttgaattg tactttttag aataatgact cactgagttg
     6121 ttgagagagt tgagggaact catagatatg ttcacagttg tttcgtgaat tcggaataca
     6181 gaatccgaat tcaaagtcaa aacacttcag aaggcgatct ttgaagaagt gacgttcgat
     6241 cattcggaaa tgatggattg ggatgtctcc tactttgaat tccacagttg ctccgaccgt
     6301 tttgagtttc aaaaagtttg gagcaaatct ataacgcacg tatcttgccg actcctgtgg
     6361 cgactcatca ttctcctgat cgttctccgg tttggcgatc tcgaaaagca cttgctcagt
     6421 gtccagatca cggatttgga acttggtgaa ctcgatgtta tagatgttcg cagatgggga
     6481 gcataagaat cctaaattta tgttttaaac tgaaatccaa agggagcaag ataccttgag
     6541 tgattcccgg aagtgctaaa acgtcgttcg gagtgatttg agctttcttc gcaagctccg
     6601 attccgttgt gattccttgt tcggtgcttg gtggtggccg tggcatctgg aaatatggaa
     6661 aagttcaaca aaaagaaaag agaaaagaat gaaatcggat atcaagagtt agttgagcgg
     6721 tttctctagt tttctgagtc tcacctgcga cgggaaggtc gccgagccgg gtggaatcga
     6781 tcgttgttgc tcggctttca tatcggtttg gttggaagcg gctgaaaacg gaaagaagtg
     6841 gaagaaggaa aagagtgtgg tgtgacagga aaatggtaat tagagggtgc caaataacca
     6901 gctatatttt gttttttttt gaaaacattt ttaaaaagaa aaatacgata atgatatcag
     6961 atggatttcc ggaaaactgg tatgaaaaat ttcaaccttt ttgagtacat gtaatcaaaa
     7021 tacactttgt aaattatcat ttttattgaa actccaccat ttttctattt ataacgctaa
     7081 taatttgaaa aagaaaccta ttgcgaaccg cggggtgaat cccaaaaacg aatgcgtttt
     7141 ggtggagtga ttgattcgaa tcgaagaaga aaaagaagaa gacgtggaat agagagctca
     7201 ctcttaaccg agcagcacac accgacagaa aaaaaaatga aatgaatgag ggtcttcttc
     7261 ttcttcttct tcgaatgatt gacagaaatg ggaaaaagag gaagattgag aagggaaaaa
     7321 ggaaggagaa aagaagcaga agaagacgtc agagaggaga ggaacgagcg gaaaagcagc
     7381 gggcgcaagt catagaagta gcagagctgg ggagaagaag acactatcca agaaaggaat
     7441 gacgagagag tatgcaaagg ggtatagggt gcagacagaa taggaacaga ataacagatg
     7501 atgagccaag aagagttgaa aagggcgatg aatttgtcat gtaacttaat ttgggtcaat
     7561 ttgagcatga tgaattgaaa tcatcccttg ttgggagtta ataaccggtt tgttatcaga
     7621 aaccctgtaa tagaagggcg ccctaacttt gagccaattc atcccggttt ctgtcaaata
     7681 tatcaaaaag tggtcaactg acaaattgtt tttgatatta taataaacat tttatccgtt
     7741 aacaattttc gaatactttt tacaaggact tggataaatt ggctcaaaga cgaagtgagt
     7801 ttgctaccat catggaatca ggaggacatt tcgacaatgt catataatgt ttgaaaaatg
     7861 attttaacaa gggtctcatt cgagataagc aatgctctga gcaagtggct cgtcgtcttc
     7921 aagaatctct tgttgatatt catttggatc ctgttttgca cgaagcatgt gccatggata
     7981 ttcagagttt ttgcaaaaaa gttccgccag gtcaaacaat ctttttaatt tttatatgat
     8041 acttcgggtg tatttttctt ctttttcctg ttcattttcg cttttttctc acttttttgt
     8101 aacaaaatct ctcacttttc tgtaagtctt aacaaaagtt caattggctc gatcaagtaa
     8161 aatttcggcc gtatgaacgt tttcaaaggt tttcattttt ttttaagttc attcacaata
     8221 tcatgaatct ctcgtccttt ggtttccggt aagtagcgaa ataatataag catacttata
     8281 aatgaaggaa ttgaatatag aataaggaaa gcataactac caataacgga atatagaggt
     8341 aatactgtaa atgtactgat aaccaccatt acagtattca ttgaatatgc cacagattga
     8401 gcaatactac gatgtttttg aggtaccagt tcagaggata taaaccacga aattggacca
     8461 actccagttc tgaaatttca ttattcgatt aaatcaaaat atatttgaaa aacctcaccc
     8521 ataagtataa ccgtagatga tcaatgcaac taggcatccg tatttcatcg ggtcaatcag
     8581 aatacgtatt tttgcgaaga atacgaatgc aattagagct aaattattga gaaatgtgaa
     8641 gagaaggaga agcttgcgtc gaccaactct gtaaacttat tcatacaatt ttgtcttcaa
     8701 attttgaacg tttaaaacta tcttattcaa tgagttagcg ggtttttata gtgagttagt
     8761 tcacgattat ctaataaaac tatgtccaaa catcctaact gcgaccgtat tctcactaac
     8821 tcgtcttaaa aacgttgaga ttattttaaa cccttcttca gtacaactac agtactgatg
     8881 cacaaataca ataataataa tacattcaga catactctga acaatgtaac ttgtttttgt
     8941 tatactgtgg taacatttat tgaaatgaaa tataacctta cttttcaatc acaattccac
     9001 cagtaattgt tccagaaaca taagctaaac ccattgcagt tgtactccat tcagcaattt
     9061 ctgactcaac atttgcattt tctagaaaga aagtagagga taaaagaaga gcccataatc
     9121 cgacagggcc c
//
