LOCUS       Exported                6726 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Optimized enzyme for genome editing in C. elegans.
ACCESSION   .
VERSION     .
KEYWORDS    pMDJ20 - pEXP [Psmu-1 | Cre Recombinase (PATCs) | smu-1 UTR]
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6726)
  AUTHORS   Aljohani MD, El Mouridi S, Priyadarshini M, Vargas-Velazquez AM, 
            Frokjaer-Jensen C
  TITLE     Engineering rules that minimize germline silencing of transgenes in 
            simple extrachromosomal arrays in C. elegans.
  JOURNAL   Nat Commun. 2020 Dec 9;11(1):6300. doi: 10.1038/s41467-020-19898-0.
  PUBMED    33298957
REFERENCE   2  (bases 1 to 6726)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6726
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     1..17
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1..17
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    37..57
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    1050..1074
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3322..3346)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4339..4359)
                     /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(4367..4384)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4367..4383)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4376..4398)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4711..4733
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4771..4789)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4857..4961
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4962..5822
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGSDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5180..5199)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5993..6581
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6482..6501
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6708..4
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 caggaaacag ctatgaccat gattacgcca agctatcaac tttgtataga aaagttgaaa
       61 tttttcaggc gggaagtggc ttcggtgcgt tcaacaaacg cgcttttgac tcgctagccg
      121 gatccggctt cagcggtttt gacaaaagag ctttcgattc actggctgga caaggtttta
      181 ctggatttga gaaacgcgcc tttgatacag tggtacggtt ttcggtggaa attccagttt
      241 ttcgaatttt caataatttt cagagcacgt caggctttga cgatttcaaa ttgtgaagag
      301 ctcactgata tttaaaattt ggttcaattc aagcacatat ttttgaggta agcaactgat
      361 tttaatgttg ttaatcaaat gaaaagtgaa atttttggaa taattttttt ttgttttcag
      421 gtagctccaa cgaacgacac acacaagccc agtcccccag tattcctgac gacagcccct
      481 gctcacacac tttttattat aattcctttt ttttcaaact tttttttcaa attttatact
      541 ttaatctcaa aaaacctatt catacgaaca caagtaccgg tgtgcgtgtg tgagaataag
      601 aatttatttt aaaaaaataa agaatgtcgt cctgtttttt catatttatt gaagaaaatg
      661 gagctggaaa aaaatcaatt gtaaaaaaca gaaaaacgtg ctgataaaat gcattgtacg
      721 caaactgtgg attttgagaa ctaacgtagt caagcaaaaa tctctggaat tttatcgatt
      781 tttaacggat aaactgttgt tttttgtaac tgtgcatgaa aaaaaatctc aggcaaaaat
      841 attttctgaa aaaaaaagag tttaaacaca aatgcaagtc cacaaaaaac tccaatttga
      901 aattcgctcc aaaaacgcaa aacacgccaa gcgctcgtgc tggcgcattt gccgatcgct
      961 tccaaaaagg ctggcgcact tctcgtgtcc gaggatattc gaattttaca cgtcattttc
     1021 agccgttttt caccgaattt tcgctgaaaa caagtttgta caaaaaagca ggctaaaaat
     1081 gtccaacctc ctcaccgtcc accaaaacct cccagccctc ccagtcgacg ccacctccga
     1141 cgaggtccgt aaggtatttt cctgcatttt tcaactggga aaatgaaaga aaatcgataa
     1201 tttcagaacc tcatggacat gttccgtgac cgtcaagcct tctccgagca cacctggaag
     1261 atgctcctct ccgtctgccg ttcctgggcc gcctggtgca agctcaacaa ccgtaagtgg
     1321 ttcccagccg agccaggtaa gttttcttat gggaaagaag gaaaaaaccg agattttact
     1381 tgaaaaattg aatttttcgc gggattttca ccaaaaattg ttgaatattc attatttcac
     1441 gctgtaaaac aaaaaaaaaa aaaatcaaaa actacgttga aatcgcgttt ttaagcgaat
     1501 tttcttcaga attgccagat tttaacccca aattttgcag tttttaaata aaatttcacc
     1561 ttttcggctc aaattgtaga ttttcctgaa aatttagtac aaaaaacaat ttcctcgtaa
     1621 atttttcaaa tagattttca gaggacgtcc gtgactacct cctctacctc caagcccgtg
     1681 gactcgccgt caagaccatc caacaacacc tcggacaact caacatgctc caccgtcgtt
     1741 ccggactccc acgtccatcc gactccaacg ccgtctccct cgtcatgcgt cgtatccgta
     1801 aggagaacgt cgacgccgga gagcgtgcca agcaagccct cgccttcgag cgtaccgact
     1861 tcgaccaagt ccgttccctc atggagaact ccgaccgttg ccaaggtaag ttatgtctac
     1921 ctgcctgcct accgcctaaa ttttttgtga agtttcttca aaaaatccag aaaaaaaaac
     1981 aattttcata cgattttttc ccttaaaatt gtgaattttc atgcttttta gccccaaaag
     2041 tcattatttg agaaaaaatt tcatacaaaa aagttttgag aaatacacaa ttttttaaat
     2101 gtaattttca aattttcaat tttcaactag aaaattcaca aaacttgtaa attttggacc
     2161 aaaacattta tacaattact ttttttgaat ctaataacta caataactac aaatttaaca
     2221 ttttcagaca tccgtaacct cgccttcctc ggaatcgcct acaacaccct cctccgtatc
     2281 gccgagatcg cccgtatccg tgtcaaggac atctcccgta ccgacggagg acgtatgctc
     2341 atccacatcg gacgtaccaa gaccctcgtc tccaccgccg gagtcgagaa ggccctctcc
     2401 ctcggagtca ccaagctcgt cgagcgttgg atctccgtct ccggagtcgc cgacgaccca
     2461 aacaactacc tcttctgccg tgtccgtaag gtaagttctt tttttgaaaa gtgcagtttg
     2521 tagtctaatt ttcatttatt ttctttttaa aaaacgcatc aatttttaaa tattttggga
     2581 caaaatccga aaactgtact aatttgtagt ttgtaaaatt aaaaaaaaaa cgcaaaaaaa
     2641 tgttttcaaa aatgcttaga ataaaaataa aatttctaaa aaattgaaca aaataaacat
     2701 ttaaaaattc aaaaagtttg aaaaaatgca aagtttttta atgcaaaaaa tatttaatgt
     2761 ttaaacaaat ttaaaaaatg ttgtaagaat atgtttagag acagttttta aatgttaaat
     2821 tgcaaaaaaa aaaagcaaaa aaaaatttga aaaatgtaac agaaaatttt aattaaaaaa
     2881 gtcgtgtttt tttcaaacct gcaattcttt gtaattttat ttataaagag aaacaaatca
     2941 aattgtatat tacgaaaaac gatactaaaa ttcgaaaatt tgcggttttt tgcgtaaaaa
     3001 atacggttcc gtaattttca gaacggagtc gccgccccat ccgccacctc ccaactctcc
     3061 acccgtgccc tcgagggaat cttcgaggcc acccaccgtc tcatctacgg agccaaggac
     3121 gactccggac aacgttacct cgcctggtcc ggacactccg cccgtgtcgg agccgcccgt
     3181 gacatggccc gtgccggagt ctccatccca gagatcatgc aagccggagg atggaccaac
     3241 gtcaacatcg tcatgaacta catccgtaac ctcgactccg agacgggagc catggtccgt
     3301 ctcctcgagg acggagacta gacccagctt tcttgtacaa agtggattaa ttttccccag
     3361 atttccccac gatttttttt gacattttta tgaaaatttt ctctaaaagt tacccctatt
     3421 ttgtttagac tgatctctta ttttatccaa tgtttcaaat ttttgaaatt gctcgaaaat
     3481 tccaattttc acatattttt gttgattttt tcgccatttt cccctgataa attaacttat
     3541 ttttgcaatt ttttctttcg aaaaaatatt ggtttctcat caccatttat cctttttttt
     3601 atggagttaa aaattgttaa aaaaatttaa ttttttttct aattttagag tcaaaacggg
     3661 ttcgaatcta gagcgtagct gtacgatttt ttggcgattt ttgcggtttt ctggtgagtt
     3721 tttttaaagt tatttttttg aaaaaaattc ttgaaaataa aaggaaattg ttttaaaaaa
     3781 aaatttcaaa gaaaaaattt tgaaaaaatt gaaacatttt ttttattttg accaaaaata
     3841 ttgaaaatta attaacattt ttttttggaa agttaatttt taatacaaat tttttaaatt
     3901 tattttacaa aaaaaattga atttctgtga aaaatcagtt ttccagtaca taatgatttg
     3961 accaaaaaaa attgaaaaaa atgatttttt aaaattaaaa ataccataaa atgctgaaaa
     4021 ttcagaaaaa taagtaaatt ttaaaaaatt ctggaaaaac atctaaaaac cccccaaata
     4081 taatattttg gaccctttta ttttctgctc gacatctcaa atatcctcca atttttcgct
     4141 tttttgatta attgaaagca tagtgggggc aggtgttata ggacaccctt ccctccactt
     4201 ttttggagca tttttcaaag ctcttccgag agcagaagct tctcaatttt tccaccccaa
     4261 aagtttgaaa ttctgccaaa aaatctcgca ttttaacaat ttttcgagat gtttcacgtc
     4321 cttttcattg cgatttccca actttattat acatagttga taattcactg gccgtcgttt
     4381 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     4441 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     4501 tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg
     4561 gtatttcaca ccgcatatgg tgcactctca gtgcaatctg ctctgatgcc gcatagttaa
     4621 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     4681 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     4741 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     4801 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     4861 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4921 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     4981 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     5041 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     5101 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     5161 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     5221 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     5281 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5341 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5401 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5461 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5521 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5581 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     5641 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     5701 tggggtcaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     5761 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5821 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     5881 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5941 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6001 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6061 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6121 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6181 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6241 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6301 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6361 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6421 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6481 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6541 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6601 ttttacggtt cctggtcttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     6661 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccagc ggataacaat
     6721 ttcaca
//
