LOCUS       sequence-389078-        5018 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes G-418 resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5018)
  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 5018)
  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: pZCS38 (Prsp-27::NeoR::unc-54 3' UTR)
FEATURES             Location/Qualifiers
     source          1..5018
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      572..1160
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1061..1080
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1314..1331
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1448..1469
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1484..1489,1490..1507,1508..1514)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1522..1538
                     /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     1527..1549
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1546..1562
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1546..1562
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1612..1628)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2450..3244
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2504..2523)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3114..3133
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3964..3980)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3973..3995)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4189..4208)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4308..4330
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4368..4386)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4454..4558
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4559..4627,4628..5018,1..401)
                     /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(4777..4796)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
       61 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      121 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      181 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      241 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      301 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
      361 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
      421 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
      481 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
      541 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
      601 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
      661 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
      721 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
      781 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
      841 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
      901 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
      961 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1021 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1081 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1141 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1201 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1261 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1321 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     1381 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     1441 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     1501 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     1561 accatgatta cgccaagctt gcatgcctgc aggtcgactc tagaggatcc cactggccgt
     1621 cgttttacag gccggatggg ccatggcacc ttaacatttc taaattttag tttaataaaa
     1681 aaattttaaa ataatacgag ttttgagact aactttcaat aaaaaccaaa aaattgcttc
     1741 aaaaaagagc tcgcgccgat ttacgataaa cacagcgcgc tacgacactc cgaaaaccgc
     1801 gccgcaccat tacattctcc gccccaccgc acggtttgcg gaaggcgatg aaattgctgg
     1861 gcctaaaatt ttaacccttt ttttcccgtt tttcaactgt tttcccgatt ttttcgatat
     1921 ttttttcagg tttttagtta gaaaagtata taaaaattgc attatttgta gttttcgctc
     1981 gtaatttgtg atagaaaacg gccaatttgc ctagatttgt gattctacca agtggaattt
     2041 ttgggaaaat atcaatattc cacgattttt attcaaatca taacgtttcg aaattatcag
     2101 cttaaataaa actaaaataa ataataaaat actaaaaacc ctgaatttct cggttctcgg
     2161 ttcagcttga gttcgttcta tacacgttat cgatttttgt catttcgttt aatcatgggt
     2221 tttgattcat tttaaatatg aaatttgatt tattcctcgt ttcgaagttg gtttgtcttg
     2281 tccggccggt aaacaaatgt tattaaaaaa aataataaat ttcgaaattt ttcgtaatgt
     2341 tccgcaatcg tgtttttccg tttatctcgt cttcccttcc ttcaaatata ctatttattt
     2401 tctcaattca catattttca gccagccctg ctcaacaagt ggaataaaaa tgattgaaca
     2461 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     2521 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     2581 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     2641 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     2701 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     2761 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     2821 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     2881 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     2941 gctcgcacca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     3001 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     3061 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     3121 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     3181 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     3241 ctgagtccaa ttactcttca acatccctac atgctctttc tccctgtgct cccaccccct
     3301 atttttgtta ttatcaaaaa acttctctta atttctttgt tttttagctt cttttaagcc
     3361 acctctaaca atgaaattgt gtagattcaa aaatagaatt aattcgtaat aaaaagtcga
     3421 aaaaaattgt gctccctccc cccattaata ataattctat cccaaaatct acacaatgtt
     3481 ctgtgtacac ttcttatgtt ttttacttct gataaatttt tttgaaacat catagaaaaa
     3541 accgcacaca aaatacctta tcatatgtta cgtttcagtt tatgaccgca atttttattt
     3601 cttcgcacgt ctgggcctct catgacgtca aatcatgctc atcgtgaaaa agttttggag
     3661 tatttttgga atttttcaat caagtgaaag tttatgaaat taattttcct gcttttgctt
     3721 tttggggttt cccctattgt ttgtcaagat ttcgaggacg gcgtttttct tgctaaaatc
     3781 acaagtattg atgagcacga tgcaagaaag atcggaagaa ggtttgggtt tgaggctcag
     3841 tggaaggtga gtagaagttg ataatttgaa agtggagtag tgtctatggg gtttttgcct
     3901 taaatgacag aatacattcc caatatacca aacataactg tttcgggtac cgagctcgaa
     3961 ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     4021 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     4081 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct
     4141 ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc
     4201 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     4261 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     4321 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     4381 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     4441 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4501 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     4561 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     4621 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     4681 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     4741 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     4801 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     4861 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     4921 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     4981 aggaccgaag gagctaaccg cttttttgca caacatgg
//
