LOCUS       sequence-218958-        5586 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Enhanced RNAi vektor for C. elegans. Contains two T7 polymerase 
            termination sequences adjacent to the T7 promoter sites and a lacZ 
            coding sequence..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5586)
  AUTHORS   Sturm A, Saskoi E, Tibor K, Weinhardt N, Vellai T
  TITLE     Highly efficient RNAi and Cas9-based auto-cloning systems for C. 
            elegans research.
  JOURNAL   Nucleic Acids Res. 2018 Jun 19. pii: 5040046. doi: 
            10.1093/nar/gky516.
  PUBMED    29924347
REFERENCE   2  (bases 1 to 5586)
  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"; journalName: "Nucleic 
            Acids Res. 2018 Jun 19. pii: 5040046. doi: 10.1093/nar/gky516."
COMMENT     Sequence Label: T777T
FEATURES             Location/Qualifiers
     source          1..5586
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(2..49)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        50..68
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     93..109
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     94..110
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             199..3246
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /translation="VVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS
                     LNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNVTYPITVNPP
                     FVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDSRLPSEF
                     DLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISDFHVATR
                     FNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERGGYADRV
                     TLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLLLLNGKP
                     LLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTLCDRYGL
                     YVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGNESGHGA
                     NHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWSIKKWLS
                     LPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKYDENGNP
                     WSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIEVTSEYL
                     FRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWLTVRVVQ
                     PNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKRWQFNRQ
                     SGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAGHYQAEA
                     ALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVASDTPHP
                     ARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPYVFPSENGLR
                     CGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHMGIGGDD
                     SWSPSVSAEFQLSAGRYHYQLVWCQK"
     primer_bind     complement(201..223)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        complement(3379..3397)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      3398..3445
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      complement(3507..4095)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3587..3606)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4266..5057,5058..5126))
                     /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     4889..4908
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5127..5231)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5394..5415)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctagt aatacgactc
       61 actatagggc gaattgggta ccgggccccc cctcgaggtc gacggtatcg ataagcttag
      121 tctccgaagg aacgtgtcac ggttgacagc agtcctaggt ataatactag tggactttaa
      181 gaaggagata tacatatggt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
      241 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
      301 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg ctttgcctgg
      361 tttccggcac cagaagcggt gccggaaagc tggctggagt gcgatcttcc tgaggccgat
      421 actgtcgtcg tcccctcaaa ctggcagatg cacggttacg atgcgcccat ctacaccaac
      481 gtgacctatc ccattacggt caatccgccg tttgttccca cggagaatcc gacgggttgt
      541 tactcgctca catttaatgt tgatgaaagc tggctacagg aaggccagac gcgaattatt
      601 tttgatggcg ttaactcggc gtttcatctg tggtgcaacg ggcgctgggt cggttacggc
      661 caggacagtc gtttgccgtc tgaatttgac ctgagcgcat ttttacgcgc cggagaaaac
      721 cgcctcgcgg tgatggtgct gcgctggagt gacggcagtt atctggaaga tcaggatatg
      781 tggcggatga gcggcatttt ccgtgacgtc tcgttgctgc ataaaccgac tacacaaatc
      841 agcgatttcc atgttgccac tcgctttaat gatgatttca gccgcgctgt actggaggct
      901 gaagttcaga tgtgcggcga gttgcgtgac tacctacggg taacagtttc tttatggcag
      961 ggtgaaacgc aggtcgccag cggcaccgcg cctttcggcg gtgaaattat cgatgagcgt
     1021 ggtggttatg ccgatcgcgt cacactacgt ctgaacgtcg aaaacccgaa actgtggagc
     1081 gccgaaatcc cgaatctcta tcgtgcggtg gttgaactgc acaccgccga cggcacgctg
     1141 attgaagcag aagcctgcga tgtcggtttc cgcgaggtgc ggattgaaaa tggtctgctg
     1201 ctgctgaacg gcaagccgtt gctgattcga ggcgttaacc gtcacgagca tcatcctctg
     1261 catggtcagg tcatggatga gcagacgatg gtgcaggata tcctgctgat gaagcagaac
     1321 aactttaacg ccgtgcgctg ttcgcattat ccgaaccatc cgctgtggta cacgctgtgc
     1381 gaccgctacg gcctgtatgt ggtggatgaa gccaatattg aaacccacgg catggtgcca
     1441 atgaatcgtc tgaccgatga tccgcgctgg ctaccggcga tgagcgaacg cgtaacgcga
     1501 atggtgcagc gcgatcgtaa tcacccgagt gtgatcatct ggtcgctggg gaatgaatca
     1561 ggccacggcg ctaatcacga cgcgctgtat cgctggatca aatctgtcga tccttcccgc
     1621 ccggtgcagt atgaaggcgg cggagccgac accacggcca ccgatattat ttgcccgatg
     1681 tacgcgcgcg tggatgaaga ccagcccttc ccggctgtgc cgaaatggtc catcaaaaaa
     1741 tggctttcgc tacctggaga gacgcgcccg ctgatccttt gcgaatacgc ccacgcgatg
     1801 ggtaacagtc ttggcggttt cgctaaatac tggcaggcgt ttcgtcagta tccccgttta
     1861 cagggcggct tcgtctggga ctgggtggat cagtcgctga ttaaatatga tgaaaacggc
     1921 aacccgtggt cggcttacgg cggtgatttt ggcgatacgc cgaacgatcg ccagttctgt
     1981 atgaacggtc tggtctttgc cgaccgcacg ccgcatccag cgctgacgga agcaaaacac
     2041 cagcagcagt ttttccagtt ccgtttatcc gggcaaacca tcgaagtgac cagcgaatac
     2101 ctgttccgtc atagcgataa cgagctcctg cactggatgg tggcgctgga tggtaagccg
     2161 ctggcaagcg gtgaagtgcc tctggatgtc gctccacaag gtaaacagtt gattgaactg
     2221 cctgaactac cgcagccgga gagcgccggg caactctggc tcacagtacg cgtagtgcaa
     2281 ccgaacgcga ccgcatggtc agaagccggg cacatcagcg cctggcagca gtggcgtctg
     2341 gcggaaaacc tcagtgtgac gctccccgcc gcgtcccacg ccatcccgca tctgaccacc
     2401 agcgaaatgg atttttgcat cgagctgggt aataagcgtt ggcaatttaa ccgccagtca
     2461 ggctttcttt cacagatgtg gattggcgat aaaaaacaac tgctgacgcc gctgcgcgat
     2521 cagttcaccc gtgcaccgct ggataacgac attggcgtaa gtgaagcgac ccgcattgac
     2581 cctaacgcct gggtcgaacg ctggaaggcg gcgggccatt accaggccga agcagcgttg
     2641 ttgcagtgca cggcagatac acttgctgat gcggtgctga ttacgaccgc tcacgcgtgg
     2701 cagcatcagg ggaaaacctt atttatcagc cggaaaacct accggattga tggtagtggt
     2761 caaatggcga ttaccgttga tgttgaagtg gcgagcgata caccgcatcc ggcgcggatt
     2821 ggcctgaact gccagctggc gcaggtagca gagcgggtaa actggctcgg attagggccg
     2881 caagaaaact atcccgaccg ccttactgcc gcctgttttg accgctggga tctgccattg
     2941 tcagacatgt ataccccgta cgtcttcccg agcgaaaacg gtctgcgctg cgggacgcgc
     3001 gaattgaatt atggcccaca ccagtggcgc ggcgacttcc agttcaacat cagccgctac
     3061 agtcaacagc aactgatgga aaccagccat cgccatctgc tgcacgcgga agaaggcaca
     3121 tggctgaata tcgacggttt ccatatgggg attggtggcg acgactcctg gagcccgtca
     3181 gtatcggcgg aattccagct gagcgccggt cgctaccatt accagttggt ctggtgtcaa
     3241 aaataataac ccttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttccgtgaca
     3301 cgttccttcg gagactctag agcggccgcc accgcggtgg agctcgaatt catcgatgat
     3361 atcagatctg ccggtctccc tatagtgagt cgtattacta gcataacccc ttggggcctc
     3421 taaacgggtc ttgaggggtt ttttgacatg tgagcaaaag gccagcaaaa ggccaggaac
     3481 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3541 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3601 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3661 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3721 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3781 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3841 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3901 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3961 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4021 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4081 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4141 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4201 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4261 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4321 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4381 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4441 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4501 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4561 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4621 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4681 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4741 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4801 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4861 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     4921 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4981 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5041 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5101 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5161 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5221 ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt
     5281 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     5341 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     5401 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     5461 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     5521 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     5581 agccgg
//
