LOCUS       sequence-208508-        6801 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the plant proteins TIR1 (necessary for Auxin-Inducible 
            Degradation) pan-neuronally in C. elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6801)
  AUTHORS   Kenneth G. Miller
  TITLE     An Intron-Compatible Marker for Long Distance CRISPR-Mediated Gene 
            Editing in Caenorhabditis elegans
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6801)
  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: 
            "Unpublished"
COMMENT     Sequence Label: KG#908
FEATURES             Location/Qualifiers
     source          1..6801
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(4241..4263)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4363..4382
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4460..4478)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4546..4650
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4651..4719,4720..5511)
                     /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(4869..4888)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5682..6270
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6171..6190
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6424..6441
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6558..6579
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6594..6599,6600..6617,6618..6624)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6632..6648
                     /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     6637..6659
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6656..6672
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6656..6672
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgggc tgcaggatct tcagatggga gcagtggact gtctatttga tattcttaaa
       61 atgtactcta taaaagatga aactgtaatg accaattccc tacataaata tttttaacta
      121 attggtcgga tatttgggga tcagagaagt acttgttttt gcgatattaa taatgatata
      181 tttttctagt aaaaaaatgt taagcgattt gtgaataatt aagttggctt gaaaaagatt
      241 ttttgatagc tatgtatttt gttaatttaa atcagatttt aaaactttat tattgactat
      301 ttatatatat tatatattat atattttcat ggtttaatca aaatctatta aattatttct
      361 aaaatctcta aatatcgaat tctctaaaaa gcttcatcaa caaaagcatt tcaattatct
      421 aacacaatga gaataattca agaactgaaa tagaaagaaa tctagaaaaa aaagagagtg
      481 aggtgcccta ttaaatcggt gctctcctct ctaacagaag cgcctacacg cacccaagag
      541 agctcttttt ctttcttttt tcggagcaca catacacgca tacactcctc agaagtgtat
      601 agagaaaatg ggagaagaag cgaaagaaga cgaagaagcg gttgtgatgt atgcatcggg
      661 cgtttctatc tctctccgtg agcaacgagc tagtcaaccc atcgatccat ttttgtgagc
      721 acacacagag agagactcaa acacactcac agcttcacac tgagctccaa gagctctgta
      781 gagagaagcc agagtatgct ttcctcgatt ttcctaacgt cttcttcttc tgcttttttc
      841 tttttcccaa cgagttgtag tccactaact tcaatttttc ctccagagaa agagttttag
      901 agagacagct gataaataca ctagagaaat aaaaagaatg ataaaaattt gctattttaa
      961 tcagttccct ctcgtttctc taagttttgt tgttgggttg ggaaattaaa attggaaaat
     1021 ggaaataacg tttttttctg tctaatcttg aattctccag acatttctta tttccttttt
     1081 tgttgcaagt ttttcttcct tctctaattt ttaatcaaag atatctcgct atgaactttg
     1141 aatttttgat attttgattc tataaataca taaagtaatc taaatgctct tttaaaataa
     1201 atctacagta gccctatttt caggatcccc gggattggcc aaaggaccca aaggtatgtt
     1261 tcgaatgata ctaacataac atagaacatt ttcaggagga cccttggcta gcaaaaatgc
     1321 aaaagagaat cgccttgtcg tttccagaag aagtactcga gcacgtcttc tccttcatcc
     1381 aactcgacaa ggaccgtaac tccgtctccc tcgtctgcaa gtcctggtac gagatcgagc
     1441 gttggtgccg tcgtaaggtc ttcatcggaa actgctacgc cgtctcccca gccaccgtca
     1501 tccgtcgttt cccaaaggtc cgttccgtcg agctcaaggg aaagccacac ttcgccgact
     1561 tcaacctcgt cccagacgga tggggaggat acgtctaccc atggatcgag gccatgtcct
     1621 cctcctacac ctggctcgag gagatccgtc tcaagcgtat ggtcgtcacc gacgactgcc
     1681 tcgagctcat cgccaagtcc ttcaagaact tcaaggtcct cgtcctctcc tcctgcgagg
     1741 gattctccac cgacggactc gccgccatcg ccgccacctg ccgtaacctc aaggtaagtt
     1801 taaacatata tatactaact aaccctgatt atttaaattt tcaggagctc gacctccgtg
     1861 agtccgacgt cgacgaggtc tccggacact ggctctccca cttcccagac acctacacct
     1921 ccctcgtctc cctcaacatc tcctgcctcg cctccgaggt ctccttctcc gccctcgagc
     1981 gtctcgtcac ccgttgccca aacctcaagt ccctcaagct caaccgtgcc gtcccactcg
     2041 agaagctcgc caccctcctc caacgtgccc cacaactcga ggagctcgga accggaggat
     2101 acaccgccga ggtccgtcca gacgtctact ccggactctc cgtcgccctc tccggatgca
     2161 aggtaagttt aaacagttcg gtactaacta accatacata tttaaatttt caggagctcc
     2221 gttgcctctc cggattctgg gacgccgtcc cagcctacct cccagccgtc tactccgtct
     2281 gctcccgtct caccaccctc aacctctcct acgccaccgt ccaatcctac gacctcgtca
     2341 agctcctctg ccaatgccca aagctccaac gtctctgggt cctcgactac atcgaggacg
     2401 ccggactcga ggtcctcgcc tccacctgca aggacctccg tgagctccgt gtcttcccat
     2461 ccgagccatt cgtcatggag ccaaacgtcg ccctcaccga gcaaggactc gtctccgtct
     2521 ccatgggatg cccaaagctc gagtccgtcc tctacttctg ccgtcaaatg accaacgccg
     2581 ccctcatcac catcgcccgt aaccgtccaa acatgacccg tttccgtctc tgcatcatcg
     2641 agccaaaggc cccagactac ctcaccctcg agccactcga catcggattc ggagccatcg
     2701 tcgagcactg caaggacctc cgtcgtctct ccctctccgg actcctcacc gacaaggtct
     2761 tcgagtacat cggaacctac gccaagaaga tggagatgct ctccgtcgcc ttcgccggag
     2821 actccgacct cggactccac cacgtcctct ccggatgcga ctccctccgt aagctcgaga
     2881 tccgtgactg cccattcgga gacaaggccc tcctcgccaa cgcctccaag ctcgagacca
     2941 tgcgttccct ctggatgtcc tcctgctccg tctccttcgg agcctgcaag ctcctcggac
     3001 aaaagatgcc aaagctcaac gtcgaggtca tcgacgagcg tggagcccca gactcccgtc
     3061 cagagtcctg cccagtcgag cgtgtcttca tctaccgtac cgtcgccgga ccacgtttcg
     3121 acatgccagg attcgtctgg aacatggacc aagactccac catgcgtttc tcccgtcaaa
     3181 tcatcaccac caacggactc taaatggtac cggtcagata tcactcgagg agctcccgag
     3241 atcttcagcg ctcagcattc gtagaattgt aagtttaaac tttctcatac taactaacgc
     3301 gctctattta aattttcaga attccaactg agcgccggtc gctaccatta ccaacttgtc
     3361 tggtgtcaaa aataataggg gccgctgtca tcagagtaag tttaaactga gttctactaa
     3421 ctaacgagta atatttaaat tttcagcatc tcgcgcccgt gcctctgact tctaagtcca
     3481 attactcttc aacatcccta catgctcttt ctccctgtgc tcccaccccc tatttttgtt
     3541 attatcaaaa aaacttcttc ttaatttctt tgttttttag cttcttttaa gtcacctcta
     3601 acaatgaaat tgtgtagatt caaaaataga attaattcgt aataaaaagt cgaaaaaaat
     3661 tgtgctccct ccccccatta ataataattc tatcccaaaa tctacacaat gttctgtgta
     3721 cacttcttat gtttttttta cttctgataa attttttttg aaacatcata gaaaaaaccg
     3781 cacacaaaat accttatcat atgttacgtt tcagtttatg accgcaattt ttatttcttc
     3841 gcacgtctgg gcctctcatg acgtcaaatc atgctcatcg tgaaaaagtt ttggagtatt
     3901 tttggaattt ttcaatcaag tgaaagttta tgaaattaat tttcctgctt ttgctttttg
     3961 ggggtttccc ctattgtttg tcaagagttt cgaggacggc gtttttcttg ctaaaatcac
     4021 aagtattgat gagcacgatg caagaaagat cggaagaagg tttgggtttg aggctcagtg
     4081 gaaggtgagt agaagttgat aatttgaaag tggagtagtg tctatggggt ttttgcctta
     4141 aatgacagaa tacattccca atataccaaa cataactgtt tcctactagt cggccgtacg
     4201 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     4261 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     4321 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     4381 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     4441 gcatcaggcg gccttaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     4501 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4561 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4621 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     4681 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     4741 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     4801 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     4861 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     4921 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     4981 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5041 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5101 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5161 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5221 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5281 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5341 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5401 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5461 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     5521 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5581 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5641 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5701 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5761 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5821 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5881 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5941 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6001 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6061 ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6121 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6181 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6241 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6301 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6361 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     6421 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     6481 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     6541 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     6601 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     6661 aacagctatg accatgatta cgccaagctg taagtttaaa catgatctta ctaactaact
     6721 attctcattt aaattttcag agcttaaaaa tggctgaaat cactcacaac gatggatacg
     6781 ctaacaactt ggaaatgaaa t
//
