LOCUS       sequence-60711-e        6589 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6589)
  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 6589)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pCFJ1259
FEATURES             Location/Qualifiers
     source          1..6589
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..349)
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     CDS             2294..2305
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     2310..2327
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2311..2327
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2335..2458
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2483..2488,2489..2506,2507..2513)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2567..3226
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2633..2652)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3568..3873
                     /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     3752..3771
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3914..4037)
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4057..4073)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4057..4073)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(4482..5070)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4562..4581)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5241..6032,6033..6101))
                     /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     5864..5883
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6102..6206)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6274..6292
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6330..6352)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6452..6471
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cctcttcgct attacgccag cttgagactt ttttcttggc ggcacaataa agtttcctct
       61 ttttgttggg tcactcaacg aacacaatac aaatatttga gcgctcgcat ttttctcaac
      121 ctttcgggtt taaagcactt caaaatggaa ttgaagagag attgtgaaag tacatcagac
      181 agaatcacat tttgatagca ttcacttcac tcagatgcaa acttttcagt gactataaaa
      241 agtcataaaa taataacatt aaaaaagcaa ataaattagc gagagaattt tttgacaaaa
      301 agaaagaaga gtgatagaga agaagggaat gcttgaaagg attttgcatt tatctcatgg
      361 attttcaaca cttggatcaa tatgagcagt tgagttgaga caattaaatc gtacagcagc
      421 gcacaaagtt ggtctcaggc cagctgtttg aactaattcc atcaacggat ggatcgcacc
      481 acgagactca taataattca gcagcaagga gaacagtgca gagttcaatg ataagagcat
      541 attctggcat gggtcaatgt aaaggacatg ggcgaaaatc tgtgcaagaa atatcaagcc
      601 aatcaaaccc ttgcaaacaa gaagcaggat agtttttgac gagacagaaa atcttccttg
      661 cttaatttca aatgggctaa aaaaaattaa ccacaatgaa gcaattgatc caataagagc
      721 aataacatgc gcaaaccgat ttacccatac attttcaggc agatcaagat acatgtagat
      781 tttgaaaatc tttacaagag caagtaaaaa tccgactatg atgttccaca aaaatatgaa
      841 aatcatcgaa aattttaagt tttggaaatc aaagcgcata ttttttctac cggtaccaat
      901 acgactcact atagggcacc cgggtgtaca gcggccgcga atcgatagga tccgatgagg
      961 attttcgaag ttttttagat gcactagaac aaagcgtgtt ggcttcctct gagcccgctt
     1021 tccttatata cccgcattct gcagccttac agaatgttct agaaggtcct agatgcattc
     1081 gtttgaaaat actcccggtg ggtgcaaaga gacgcagacg gaaaatgtat ctgggtctct
     1141 ttattgtgta cactactttt ccatgtaccg aatgtgagtc gccctccttt tgcaacaagc
     1201 agctcgaatg ttctagaaaa aggtggaaaa tagtataaat accgttgaaa ataaataccg
     1261 aacaacattt gctctaattg tgaaattaga aatcttcaaa ctataatcat gtcactttac
     1321 cactatttcc gtccagctca acgttccgtt tttggtgatc ctctagagtc gacctgcagg
     1381 ccgggctacg taatacgact ctgctgcaag gcgattaagt tgggtaacgc caggggggta
     1441 ccagagctca cctaggtatc tagatatcag gtgtacaagt atgaaatgtc gtttttttta
     1501 aatcaaaaaa cacgttaaat tttgtggaaa aagaaaaaat catttattca aagtatttgc
     1561 cgtcgctagc tacacatttt tcccatctct cgggcaattt gtggattcca cgccagtaga
     1621 actcatcgtc ttttgcggcg aaccattcat cgagccattt tttcacactt tcgtaagaat
     1681 cgaagcgctg ctcagcgagt gcgtgtccca tcgaagcgaa taggtggtaa tcggatgggg
     1741 ccaggtctgg tgagtaagcc gcagatccgg cacccggggt gaatcccaaa aacgaatgcg
     1801 ttttggtgga gtgattgatt cgaatcgaag aagaaaaaga agaagacgtg gaatagagag
     1861 ctcactctta accgagcagc acacaccgac agaaaaaaaa atgaaatgaa tgagggtctt
     1921 cttcttcttc ttcttcgaat gattgacaga aatgggaaaa agaggaagat tgagaaggga
     1981 aaaaggaagg agaaaagaag cagaagaaga cgtcagagag gagaggaacg agcggaaaag
     2041 cagcgggcgc aagtcataga agtagcagag ctggggagaa gaagacacta tccaagaaag
     2101 gaatgacgag agagtatgca aaggggtata gggtgcagac agaataggaa cagaataaca
     2161 gatgatgagc caagaagagt tgaaaagggc gatgaatttg tcatgtaact taatttgggt
     2221 caatttgagc atgatgaatt gaaatcatcc cttgttggga gttaataacc ggtttgttat
     2281 cagaaaccct gtaatagaag ggcgtgccat gtaaaacgac ggccagtgaa ttatcaacta
     2341 tgtataataa agttgaacga gaaacgtaaa atgatataaa tatcaatata ttaaattaga
     2401 ttttgcataa aaaacagact acataatact gtaaaacaca acatatccag tcactatggc
     2461 ggccgcatta ggcaccccag gctttacact ttatgcttcc ggctcgtata atgtgtggat
     2521 tttgagttag gatccgtcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat
     2581 cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt
     2641 tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt
     2701 aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg
     2761 cctgatgaat gctcatccgg aattccgtat ggcaatgaaa gacggtgagc tggtgatatg
     2821 ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct
     2881 ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc
     2941 gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt
     3001 ctcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa
     3061 cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat
     3121 gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct
     3181 taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaatcta gaggatccgg
     3241 cttactaaaa gccagataac agtatgcgta tttgcgcgct gatttttgcg gtataagaat
     3301 atatactgat atgtataccc gaagtatgtc aaaaagaggt atgctatgaa gcagcgtatt
     3361 acagtgacag ttgacagcga cagctatcag ttgctcaagg catatatgat gtcaatatct
     3421 ccggttcggt aagcacaacc atgcagaatg aagcccgtcg tctgcgtgcc gaacgctgga
     3481 aagcggaaaa tcaggaaggg atggctgagg tcgcccggtt tattgaaatg aacggctctt
     3541 ttgccgacga gaacaggggc tggtgaaatg cagtttaagg tttacaccta taaaagagag
     3601 agccgttatc gtctgtttgt ggatgtacag agtgatatta ttgacacgcc cgggcgacgg
     3661 atggtgatcc ccctggccag tgcacgtctg ctgtcagata aagtcccccg tgaactttac
     3721 ccggtggtgc atatcgggga tgaaagctgg cgcatgatga ccaccgatat ggccagtgtg
     3781 ccggtctccg ttatcgggga agaagtggct gatctcagcc accgcgaaaa tgacatcaaa
     3841 aacgccatta acctgatgtt ctggggaata taaatgtcag gctcccttat acacagccag
     3901 tctgcaggtc gaccatagtg actggatatg ttgtgtttta cagtattatg tagtctgttt
     3961 tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt ctcgttcaac
     4021 ttttctatac aaagttgata gcttggcgta atcatggtca tagctgtttc ctggatccgg
     4081 ccgcagctgt tttcttcaaa tgaaaacaga aaattaatac tgtccgcgtt tgctctttat
     4141 tcggcacgaa actcgacatg ttgactgcac tgagagtaaa caattatgac gctcaattcg
     4201 cgccaaacta tggtggttcg acagtcaagg ttgacacttc acaaggtcaa agttttatga
     4261 caatcgataa atatttacgt ttgcgagaca tctatatgtt cgaaacgaca tttcatactt
     4321 gtacacctga tagaattcct cgagacgtac ggtgcggtat cagctcactc aaaggcggta
     4381 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4441 caaaagacca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4501 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4561 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4621 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4681 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4741 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4801 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4861 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4921 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4981 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5041 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5101 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5161 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5221 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5281 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5341 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5401 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5461 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5521 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5581 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5641 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5701 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5761 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5821 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5881 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5941 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6001 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6061 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6121 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6181 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     6241 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     6301 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     6361 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     6421 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     6481 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     6541 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcggg
//
