LOCUS       sequence-307911-        4339 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  (eft-3p::wrmScarlet::tbb-2 3'UTR in pUC19) Experimentally used to 
            mark formation of transgenic arrays for C. elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4339)
  AUTHORS   Stevenson ZC, Moerdyk-Schauwecker MJ, Jamison B, Phillips PC
  TITLE     Rapid Self-Selecting and Clone-Free Integration of Transgenes into 
            Engineered CRISPR Safe Harbor Locations in Caenorhabditis elegans.
  JOURNAL   G3 (Bethesda). 2020 Aug 19. pii: g3.120.401400. doi: 
            10.1534/g3.120.401400.
  PUBMED    32816924
REFERENCE   2  (bases 1 to 4339)
  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: "G3 
            (Bethesda). 2020 Aug 19. pii: g3.120.401400. doi: 
            10.1534/g3.120.401400."
COMMENT     Sequence Label: pZCS16 (Peft-3::wrmScarlet::tbb-2 3'UTR in pUC19)
FEATURES             Location/Qualifiers
     source          1..4339
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      83..671
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     572..591
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     825..842
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    959..980
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(995..1000,1001..1018,1019..1025)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1033..1049
                     /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     1038..1060
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1057..1073
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1057..1073
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1123..1722
                     /gene="eft-3"
                     /label=eft-3 promoter
                     /note="promoter for the C. elegans elongation factor 
                     1-alpha gene (Mitrovich and Anderson, 2000)"
     terminator      2449..2772
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     primer_bind     complement(2796..2813)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2796..2812)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2805..2827)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3021..3040)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3140..3162
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3200..3218)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3286..3390
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3391..3459,3460..4251)
                     /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(3609..3628)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
       61 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
      121 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
      181 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
      241 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
      301 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
      361 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
      421 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
      481 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
      541 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
      601 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
      661 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
      721 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
      781 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
      841 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
      901 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
      961 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     1021 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     1081 acgccaagct tgcatgcctg caggtcgact ctagaggatc ccgcaccttt ggtcttttat
     1141 tgtcaacttc cattggttct tccattgttt ctgttaaatt aatgaatttt tcataaaata
     1201 aagacattat acaatataaa aatgaagaat ttattgaaaa taaactgcca gagagaaaaa
     1261 gtatgcaaca ctcccgccga gagtgtttga aatggtgtac ggtacatttt cgtgctagga
     1321 gttagatgtg caggcagcaa cgagaggggg agagattttt ttgggccttg tgaaattaac
     1381 gtgagttttc tggtcatctg actaatcatg ttggtttttt gttggtttat tttgttttta
     1441 tctttgtttt tatccagatt aggaaattta aattttatga atttataatg aggtcaaaca
     1501 ttcagtccca gcgtttttcc tgttctcact gtttagtcga atttttattt taggctttca
     1561 acaaatgttc taactgtctt atttgtgacc tcacttttta tattttttta atttttaaaa
     1621 atattagaag tttctaggat aattttttcg acttttattc tctctaccgt ccgcactctt
     1681 cttactttta aattaaattg tttttttttc agttgggaaa cactttgctc tagaaaaaat
     1741 ggtcagcaag ggagaggcag ttatcaagga gttcatgcgt ttcaaggtcc acatggaggg
     1801 atccatgaac ggacacgagt tcgagatcga gggagaggga gagggacgtc catacgaggg
     1861 aacccaaacc gccaagctca aggtcaccaa gggaggacca ctcccattct cctgggacat
     1921 cctctcccca caattcatgt acggatcccg tgccttcacc aagcacccag ccgacatccc
     1981 agactactac aagcaatcct tcccagaggg attcaagtgg gagcgtgtca tgaacttcga
     2041 ggacggagga gccgtcaccg tcacccaaga cacctccctc gaggacggaa ccctcatcta
     2101 caaggtcaag ctccgtggaa ccaacttccc accagacgga ccagtcatgc aaaagaagac
     2161 catgggatgg gaggcctcca ccgagcgtct ctacccagag gacggagtcc tcaagggaga
     2221 catcaagatg gccctccgtc tcaaggacgg aggacgttac ctcgccgact tcaagaccac
     2281 ctacaaggcc aagaagccag tccaaatgcc aggagcctac aacgtcgacc gtaagctcga
     2341 catcacctcc cacaacgagg actacaccgt cgtcgagcaa tacgagcgtt ccgagggacg
     2401 tcactccacc ggaggaatgg acgagctcta caagtaaact agtgataaat gcaaaatcct
     2461 ttcaagcatt cccttcttct ctatcactct tctttctttt tgtcaaaaaa ttctctcgct
     2521 aatttatttg cttttttaat gttattattt tatgactttt tatagtcact gaaaagtttg
     2581 catctgagtg aagtgaatgc tatcaaaatg tgattctgtc tgatgtactt tcacaatctc
     2641 tcttcaattc cattttgaag tgctttaaac ccgaaaggtt gagaaaaatg cgagcgctca
     2701 aatatttgta ttgtgttcgt tgagtgaccc aacaaaaaga ggaaacttta ttgtgccgcc
     2761 aagaaaaaag tctcacgggt accgagctcg aattcactgg ccgtcgtttt acaacgtcgt
     2821 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     2881 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     2941 aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     3001 cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     3061 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     3121 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     3181 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     3241 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3301 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3361 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     3421 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3481 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3541 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3601 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     3661 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3721 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3781 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3841 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     3901 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     3961 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     4021 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     4081 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     4141 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4201 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4261 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4321 taggtgaaga tcctttttg
//
