LOCUS       sequence-212553-        6795 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 6795)
  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  JOURNAL   Unpublished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 6795)
  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            "Unpublished"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..6795
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(4305..4327)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4427..4446
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4524..4542)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4610..4714
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4715..4783,4784..5575)
                     /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(4933..4952)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5746..6334
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6235..6254
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6488..6505
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6622..6643
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6658..6663,6664..6681,6682..6688)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6696..6712
                     /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     6701..6723
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6720..6736
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6720..6736
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aattttcaga gcttaaaaat ggctgaaatc actcacaacg atggatacgc taacaacttg
       61 gaaatgaaat aagcttgggc tgcaggatct tcagatggga gcagtggact gtctatttga
      121 tattcttaaa atgtactcta taaaagatga aactgtaatg accaattccc tacataaata
      181 tttttaacta attggtcgga tatttgggga tcagagaagt acttgttttt gcgatattaa
      241 taatgatata tttttctagt aaaaaaatgt taagcgattt gtgaataatt aagttggctt
      301 gaaaaagatt ttttgatagc tatgtatttt gttaatttaa atcagatttt aaaactttat
      361 tattgactat ttatatatat tatatattat atattttcat ggtttaatca aaatctatta
      421 aattatttct aaaatctcta aatatcgaat tctctaaaaa gcttcatcaa caaaagcatt
      481 tcaattatct aacacaatga gaataattca agaactgaaa tagaaagaaa tctagaaaaa
      541 aaagagagtg aggtgcccta ttaaatcggt gctctcctct ctaacagaag cgcctacacg
      601 cacccaagag agctcttttt ctttcttttt tcggagcaca catacacgca tacactcctc
      661 agaagtgtat agagaaaatg ggagaagaag cgaaagaaga cgaagaagcg gttgtgatgt
      721 atgcatcggg cgtttctatc tctctccgtg agcaacgagc tagtcaaccc atcgatccat
      781 ttttgtgagc acacacagag agagactcaa acacactcac agcttcacac tgagctccaa
      841 gagctctgta gagagaagcc agagtatgct ttcctcgatt ttcctaacgt cttcttcttc
      901 tgcttttttc tttttcccaa cgagttgtag tccactaact tcaatttttc ctccagagaa
      961 agagttttag agagacagct gataaataca ctagagaaat aaaaagaatg ataaaaattt
     1021 gctattttaa tcagttccct ctcgtttctc taagttttgt tgttgggttg ggaaattaaa
     1081 attggaaaat ggaaataacg tttttttctg tctaatcttg aattctccag acatttctta
     1141 tttccttttt tgttgcaagt ttttcttcct tctctaattt ttaatcaaag atatctcgct
     1201 atgaactttg aatttttgat attttgattc tataaataca taaagtaatc taaatgctct
     1261 tttaaaataa atctacagta gccctatttt cagggatccc cgggattggc caaaggaccc
     1321 aaaggtatgt ttcgaatgat actaacataa catagaacat tttcaggagg acccttggct
     1381 agcaaaaatg caaaagagaa tcgccttgtc gtttccagaa gaagtactcg agcacgtctt
     1441 ctccttcatc caactcgaca aggaccgtaa ctccgtctcc ctcgtctgca agtcctggta
     1501 cgagatcgag cgttggtgcc gtcgtaaggt cttcatcgga aactgctacg ccgtctcccc
     1561 agccaccgtc atccgtcgtt tcccaaaggt ccgttccgtc gagctcaagg gaaagccaca
     1621 cttcgccgac ttcaacctcg tcccagacgg atggggagga tacgtctacc catggatcga
     1681 ggccatgtcc tcctcctaca cctggctcga ggagatccgt ctcaagcgta tggtcgtcac
     1741 cgacgactgc ctcgagctca tcgccaagtc cttcaagaac ttcaaggtcc tcgtcctctc
     1801 ctcctgcgag ggattctcca ccgacggact cgccgccatc gccgccacct gccgtaacct
     1861 caaggtaagt ttaaacatat atatactaac taaccctgat tatttaaatt ttcaggagct
     1921 cgacctccgt gagtccgacg tcgacgaggt ctccggacac tggctctccc acttcccaga
     1981 cacctacacc tccctcgtct ccctcaacat ctcctgcctc gcctccgagg tctccttctc
     2041 cgccctcgag cgtctcgtca cccgttgccc aaacctcaag tccctcaagc tcaaccgtgc
     2101 cgtcccactc gagaagctcg ccaccctcct ccaacgtgcc ccacaactcg aggagctcgg
     2161 aaccggagga tacaccgccg aggtccgtcc agacgtctac tccggactct ccgtcgccct
     2221 ctccggatgc aaggtaagtt taaacagttc ggtactaact aaccatacat atttaaattt
     2281 tcaggagctc cgttgcctct ccggattctg ggacgccgtc ccagcctacc tcccagccgt
     2341 ctactccgtc tgctcccgtc tcaccaccct caacctctcc tacgccaccg tccaatccta
     2401 cgacctcgtc aagctcctct gccaatgccc aaagctccaa cgtctctggg tcctcgacta
     2461 catcgaggac gccggactcg aggtcctcgc ctccacctgc aaggacctcc gtgagctccg
     2521 tgtcttccca tccgagccat tcgtcatgga gccaaacgtc gccctcaccg agcaaggact
     2581 cgtctccgtc tccatgggat gcccaaagct cgagtccgtc ctctacttct gccgtcaaat
     2641 gaccaacgcc gccctcatca ccatcgcccg taaccgtcca aacatgaccc gtttccgtct
     2701 ctgcatcatc gagccaaagg ccccagacta cctcaccctc gagccactcg acatcggatt
     2761 cggagccatc gtcgagcact gcaaggacct ccgtcgtctc tccctctccg gactcctcac
     2821 cgacaaggtc ttcgagtaca tcggaaccta cgccaagaag atggagatgc tctccgtcgc
     2881 cttcgccgga gactccgacc tcggactcca ccacgtcctc tccggatgcg actccctccg
     2941 taagctcgag atccgtgact gcccattcgg agacaaggcc ctcctcgcca acgcctccaa
     3001 gctcgagacc atgcgttccc tctggatgtc ctcctgctcc gtctccttcg gagcctgcaa
     3061 gctcctcgga caaaagatgc caaagctcaa cgtcgaggtc atcgacgagc gtggagcccc
     3121 agactcccgt ccagagtcct gcccagtcga gcgtgtcttc atctaccgta ccgtcgccgg
     3181 accacgtttc gacatgccag gattcgtctg gaacatggac caagactcca ccatgcgttt
     3241 ctcccgtcaa atcatcacca ccaacggact ctaaatggta ccggtcagat atcactcgag
     3301 gagctcccga gatcttcagc gctcagcatt cgtagaattg taagtttaaa ctttctcata
     3361 ctaactaacg cgctctattt aaattttcag aattccaact gagcgccggt cgctaccatt
     3421 accaacttgt ctggtgtcaa aaataatagg ggccgctgtc atcagagtaa gtttaaactg
     3481 agttctacta actaacgagt aatatttaaa ttttcagcat ctcgcgcccg tgcctctgac
     3541 ttctaagtcc aattactctt caacatccct acatgctctt tctccctgtg ctcccacccc
     3601 ctatttttgt tattatcaaa aaacttctct taatttcttt gttttttagc ttcttttaag
     3661 tcacctctaa caatgaaatt gtgtagattc aaaaatagaa ttaattcgta ataaaaagtc
     3721 gaaaaaaatt gtgctccctc cccccattaa taataattct atcccaaaat ctacacaatg
     3781 ttctgtgtac acttcttatg ttttttactt ctgataaatt tttttgaaac atcatagaaa
     3841 aaaccgcaca caaaatacct tatcatatgt tacgtttcag tttatgaccg caatttttat
     3901 ttcttcgcac gtctgggcct ctcatgacgt caaatcatgc tcatcgtgaa aaagttttgg
     3961 agtatttttg gaatttttca atcaagtgaa agtttatgaa attaattttc ctgcttttgc
     4021 tttttggggt ttcccctatt gtttgtcaag atttcgagga cggcgttttt cttgctaaaa
     4081 tcacaagtat tgatgagcac gatgcaagaa agatcggaag aaggtttggg tttgaggctc
     4141 agtggaaggt gagtagaagt tgataatttg aaagtggagt agtgtctatg gggtttttgc
     4201 cttaaatgac agaatacatt cccaatatac caaacataac tgtttcctac tagtcggccg
     4261 tacgggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     4321 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     4381 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     4441 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     4501 taccgcatca ggcggcctta agggcctcgt gatacgccta tttttatagg ttaatgtcat
     4561 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4621 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4681 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     4741 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     4801 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     4861 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     4921 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     4981 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     5041 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     5101 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     5161 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     5221 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     5281 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     5341 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5401 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     5461 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5521 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     5581 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5641 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5701 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5761 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5821 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5881 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5941 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     6001 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6061 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6121 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6181 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6241 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6301 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6361 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     6421 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     6481 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     6541 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     6601 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     6661 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     6721 aggaaacagc tatgaccatg attacgccaa gctgtaagtt taaacatgat cttactaact
     6781 aactattctc attta
//
