LOCUS       sequence-389077-        3273 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes guide RNA expression targeting PX740 landing pad.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3273)
  AUTHORS   Stevenson ZC, Moerdyk-Schauwecker MJ, Banse SA, Patel DS, Lu H, 
            Phillips PC
  TITLE     High-throughput library transgenesis in Caenorhabditis elegans via 
            Transgenic Arrays Resulting in Diversity of Integrated Sequences 
            (TARDIS).
  JOURNAL   Elife. 2023 Jul 4;12:RP84831. doi: 10.7554/eLife.84831.
  PUBMED    37401921
REFERENCE   2  (bases 1 to 3273)
  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: "Elife."; 
            date: "2023-07-4"; volume: "12:RP84831. doi"; pages: " 
            10.7554/eLife.84831"
COMMENT     Sequence Label: pZCS41 (U6p::GCGAAGTGACGGTAGACCGT)
FEATURES             Location/Qualifiers
     source          1..3273
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      645..1233
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1134..1153
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1387..1404
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1521..1542
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1557..1562,1563..1580,1581..1587)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1595..1611
                     /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     1600..1622
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1619..1635
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1619..1635
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1706..1722)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1743..2092
                     /label=CeU6 promoter
                     /note="promoter for the C. elegans R07E5.16 U6 snRNA gene 
                     (Dickinson et al., 2013)"
     misc_RNA        2114..2189
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(2292..2308)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2517..2536)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2636..2658
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2696..2714)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2782..2886
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2887..2955,2956..3273,1..474)
                     /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     complement(3105..3124)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
       61 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
      121 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
      181 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
      241 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
      301 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
      361 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
      421 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
      481 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
      541 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
      601 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
      661 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
      721 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
      781 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
      841 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
      901 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
      961 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     1021 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     1081 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     1141 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     1201 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     1261 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     1321 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     1381 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     1441 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     1501 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     1561 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     1621 ggaaacagct atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga
     1681 tcccattata catagttgat aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     1741 accaaaaaaa actagcaata aaggaataaa aaactgtaca ccttaaaggc gcacactctg
     1801 ttttgcaaat tttattttta gttgtgaatt ttctgctgag acctgaaaat agcaacttta
     1861 gtactactat aatttgtcaa ccttttcaaa aaaagcatgc aatttttgag aaactcttat
     1921 aaaagctatt attaaaaaaa cacctttttt ccaaaattat tccacaaaaa atatgttatg
     1981 aaatgcctac accctctcac acacactctt tatactactc tgtcaaactc acgagatgtc
     2041 tgccgcctct tgtgttgccc ctatataaac acctcctatt gcgagatgtc ttggcgaagt
     2101 gacggtagac cgtgttttag agctagaaat agcaagttaa aataaggcta gtccgttatc
     2161 aacttgaaaa agtggcaccg agtcggtgct ttttgtgaaa tttctggcgt aatagcgaag
     2221 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tcgggtaccg
     2281 agctcgaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     2341 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     2401 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg
     2461 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca
     2521 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     2581 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     2641 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     2701 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     2761 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     2821 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     2881 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     2941 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     3001 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     3061 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     3121 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     3181 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     3241 gaattatgca gtgctgccat aaccatgagt gat
//
