LOCUS       sequence-324027-        8147 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses 3xHA-tagged TurboID in C. elegans intestine.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8147)
  AUTHORS   Branon TC, Bosch JA, Sanchez AD, Udeshi ND, Svinkina T, Carr SA, 
            Feldman JL, Perrimon N, Ting AY
  TITLE     Efficient proximity labeling in living cells and organisms with 
            TurboID.
  JOURNAL   Nat Biotechnol. 2018 Aug 20. pii: nbt.4201. doi: 10.1038/nbt.4201.
  PUBMED    30125270
REFERENCE   2  (bases 1 to 8147)
  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: "Nat 
            Biotechnol. 2018 Aug 20. pii: nbt.4201. doi: 10.1038/nbt.4201."
COMMENT     Sequence Label: 3xHA-TurboID_pAS31
FEATURES             Location/Qualifiers
     source          1..8147
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..125
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(126..194,195..986)
                     /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(344..363)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1157..1745
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1646..1665
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1899..1916
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2033..2054
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2069..2074,2075..2092,2093..2099)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2107..2123
                     /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     2112..2134
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2131..2147
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2131..2147
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2166..2186
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2168..2186
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             5582..5662
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     primer_bind     complement(7500..7519)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7501..7519)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7529..7546)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7529..7545)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7538..7560)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7687..8142
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7774..7793)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7984..8005
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
       61 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
      121 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
      181 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
      241 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
      301 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
      361 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
      421 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
      481 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
      541 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
      601 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
      661 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
      721 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
      781 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
      841 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
      901 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
      961 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     1021 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     1081 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     1141 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1201 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1261 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     1321 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     1381 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     1441 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     1501 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     1561 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     1621 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     1681 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     1741 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     1801 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     1861 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     1921 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     1981 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     2041 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     2101 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     2161 agcgcgcaat taaccctcac taaagggaac aaaagctgga gctccaccgc ggtggcggcc
     2221 gctctagaac tagtacctac tggcgcgccc taagcttaat gaagtttatt tcagatcagt
     2281 aattcgaaat gtttctactg gaatccgcca aattgtcgac aactctttta agaaaacgac
     2341 taaaatgaac ttgagtgtta gcggcgtctt caccaatacc tttagtgacg atgtcgaagt
     2401 tgactataac tgggacaata tcatcgaata ttcttacaaa tcttcggaaa cggagtataa
     2461 gaaaagagag gacgatggag accatacgga aatagctgtt agagagctcg gaattcagct
     2521 caacaatccg aatcatcagc tggagtacct ggaattctgg ataaaacctt tacgatattt
     2581 taccgaagaa gaaaggaccg aacgatggaa ttgtctggaa aaattcttca agaaggttga
     2641 gaactcgaag gttgctaaaa tctcgttcgt tggattcagg ggatataaaa ttgcaacgat
     2701 tttgcaatct ttctcaccag gaaccctgac acgactttat gtcaatgttg ctgattcgga
     2761 tatctctcaa atcatcgacg cggagcattg gatttatcta aaaaagatca gagtgctttc
     2821 gccttctccg gtttccattc cgattgatgc tattcttcac atgacacact tcgacgtttc
     2881 tgtgatccac ctttccactg atgatgtcgc aaagatcaaa aatgtgagtg aacaataaaa
     2941 agttgaaaaa acattatttt aaaataatta ctggtcgttc tcgaatccgc aagtttcaca
     3001 atggggatga tccgtaccaa catcacggac ccaatggaaa ctgcaaaaat cttcggaacg
     3061 cacattcatc gtgatgatgt ggatgttttg ggtgacatgg atattaacaa ggacgatggt
     3121 ccagaagttc atttggatga cgattggcat gaggaggagc tcgtgggggc tgtgggggcg
     3181 gaggaggagg aggacgagga ggacgaggaa gaggaggacg aggacgagga agaggggatg
     3241 gctgatgtcc cggaagtcaa tgaggccgtg cacagagtca acattaaact cggcgacgct
     3301 gacatcaccg ttgtggtgtc tccgacatac ttgaagattg caaagaatac tcgtgaaaat
     3361 tgaactttaa ttcctttctc tttttttttg actcaaaaaa taaatagtat ttcaaaggaa
     3421 tcgcatttca aactgatact taagtttttc aataggtttt ttttttcagt tttgattttt
     3481 ttttgtgctt aaagagaaga ctgttagcca ctcagccact tcagccatat cgaaacagtc
     3541 atattggctc tctttcagat cacttaaaac tccgaactat gatgacgaaa aaatgttttc
     3601 ttgaattttc attttattgt gaaattgaaa actctgcata gcccgcagtc acttgaaagc
     3661 acatttaaga tggaatcaaa tggtaagaaa gctcgtgtaa tcaggagatt ttatccgggg
     3721 tatgccgagg aatatatggt gcactcgcat tagctggcat tttttgtaat tatcggattt
     3781 ctaccttttt cttaatctag tttgttatca ttgtccatca tttacaacca aaaatttgcg
     3841 tcatttacaa ccaaaaattg atctgacaat tatcataatt cggtttttat tttcagaaga
     3901 aacgctccga tgtgatccta tagtgaatat acttttccat cctctctgaa ataaacaatt
     3961 atgaagaatg catcaatttg cattcgaaat aactgccgta ttttgtctat tggtatggct
     4021 gcagtaccat ttttcttcgc gtctttgcgc agccttcggg cgctaccaat aaggctaagc
     4081 cggggtaagt tatttcagaa ttctaatcaa ctaatctaag tctcatacat cattgtcaag
     4141 tgacgcattt ttttaatttc tatgttaact cgactttaat attcttttct ggtttcacca
     4201 gaaaaatgct ggactgaaaa attgaatttc ccgcgatttg ggaaaaaccg cagtttaagc
     4261 ttttggcatg aatacagtga acacaggaac aaaaaattga cttgttattt tgagtttata
     4321 ttaaatgtta attttgcatc tcatattttc acaaaacttg tttgcttgaa aaatgtgcaa
     4381 ataacaagtg aacatacatc aaggtttatg cagatcagca ctgatgcaac tgatagcaaa
     4441 actgataagg gtcaaaattt cagtggccag cacaaacaca tacacattcg tagttcgttt
     4501 tgtgtttcgt gtatttagca tttatttttc aattccggca atttgccgat ttgccggaaa
     4561 ttttgatttt tggcaaattg ccagtttgcc gtttgccgaa catcaatttg ccgaaatttt
     4621 ttagagggat ctttataaga cggaaacact taaaattgtg cttttttgga atttttttca
     4681 atgtttcaat agaattagct tacatttaaa attagatgta ggaacgttta taggacgctt
     4741 acaattttgc cgattaaaat cgaagttctg aacttttcaa aaaatgtgca aaattttttt
     4801 tgacaaattt gacaaatcgg ccggtttgtc gattgtccgg aatttttcag tttcggcgat
     4861 ttaccggttt gccggaaaac atcgtttgcc acccacacac acataaagag acgaaacaac
     4921 acaccatctt caaaaaattt gtgaatcttc atttccacgt ggtttgcagg gtttcccatc
     4981 caccccactc tatctctcca agttgaattt tggcaaaatt gaccacacac cgatcaaaaa
     5041 ctattgtgtg tgtgtgtatt tgtgtcttca ataattcctg tctgtgtctc gcactatcta
     5101 ttgactacaa gaacgtgatc gcagctctac gaaaccttta ctttatttta gaaatatttt
     5161 ttcatgctat tagcaaaaaa aaagtttttt tttgtctaag tacacttaaa attttccaaa
     5221 atttttaaga tttttcaaaa tgtgcgatca aagatctgac aaaatgccaa aattttgaag
     5281 aggaaattta gtcggcttcc aaaattataa gtggcagaaa ttgagcaatt gtcacttttt
     5341 gacagtaaat agaaaatttt ttgaaaaatt ttgaaacgct ttattttgtt acttggtcat
     5401 tttggcacca taggagtagt ttttaacatt ttccccactg gcgctacttc attttaaact
     5461 tttccaaaga ttttaatttt taattttaaa aaatttcata taattcgtct gcgtcttaca
     5521 gtttcaggct tcaaaaatct attacatatc ttatctttga attcagggcc ggccaaaaat
     5581 gtacccatac gacgtcccag actacgccta cccatatgat gtcccggatt acgcttaccc
     5641 atacgatgtt ccagattacg ctaaggataa caccgtccca cttaagctta tcgcccttct
     5701 tgctaacgga gaattccact ctggagagca acttggagag actcttggaa tgtcccgtgc
     5761 tgctatcaac aagcatatcc aaactcttcg tgattgggga gttgatgttt tcactgttcc
     5821 aggtaagttt aaacatatat atactaacta accctgatta tttaaatttt caggaaaggg
     5881 atactctctt ccagagccaa tcccacttct taacgctaag caaatccttg gacaacttga
     5941 tggaggatcc gtcgctgtcc ttccagttgt tgattccacc aaccaatacc ttcttgatcg
     6001 tatcggagag cttaagtctg gagacgcttg catcgctgaa taccagcagg ctggtagagg
     6061 taagtttaaa cagttcggta ctaactaacc atacatattt aaattttcag gaagtcgtgg
     6121 ccgtaaatgg tttagtcctt tcggcgccaa cctctacctt agtatgtttt ggcgtctgaa
     6181 aagaggacct gcggcgatag gcttgggtcc agtgatcggt attgttatgg ccgaggcgct
     6241 gcgaaagctg ggagctgata aggttcgtgt taagtggcca aacgatcttt accttcaaga
     6301 ccgtaagctt gctggaatcc ttgtcgagct tgctggaatc accggagacg ctgctcaaat
     6361 cgttatcgga gctggaatca acgttgctat gcgtcgtgtt gaggtaagtt taaacatgat
     6421 tttactaact aactaatctg atttaaattt tcaggagtct gttgttaacc aaggatggat
     6481 cactcttcaa gaggctggaa tcaaccttga tcgtaacacc cttgctgcca cccttatccg
     6541 tgagcttcgt gctgcccttg aacttttcga gcaagaggga cttgctccat accttccacg
     6601 ttgggagaag cttgataact tcatcaaccg cccagttaag cttatcatcg gagataagga
     6661 aatcttcgga atctctcgcg gaatcgataa gcaaggagct cttcttcttg agcaagatgg
     6721 agttattaaa ccatggatgg gaggagaaat ttcccttcgt tccgccgaga agtaacccgg
     6781 gcatctcgcg cccgtgcctc tgacttctaa gtccaattac tcttcaacat ccctacatgc
     6841 tctttctccc tgtgctccca ccccctattt ttgttattat caaaaaactt ctcttaattt
     6901 ctttgttttt tagcttcttt taagtcacct ctaacaatga aattgtgtag attcaaaaat
     6961 agaattaatt cgtaataaaa agtcgaaaaa aattgtgctc cctcccccca ttaataataa
     7021 ttctatccca aaatctacac aatgttctgt gtacacttct tatgtttttt acttctgata
     7081 aatttttttg aaacatcata gaaaaaaccg cacacaaaat accttatcat atgttacgtt
     7141 tcagtttatg accgcaattt ttatttcttc gcacgtctgg gcctctcatg acgtcaaatc
     7201 atgctcatcg tgaaaaagtt ttggagtatt tttggaattt ttcaatcaag tgaaagttta
     7261 tgaaattaat tttcctgctt ttgctttttg gggtttcccc tattgtttgt caagatttcg
     7321 aggacggcgt ttttcttgct aaaatcacaa gtattgatga gcacgatgca agaaagatcg
     7381 gaagaaggtt tgggtttgag gctcagtgga aggtgagtag aagttgataa tttgaaagtg
     7441 gagtagtgtc tatggggttt ttgccttaaa tctcgagggg gggcccggta cccaattcgc
     7501 cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg
     7561 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     7621 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     7681 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     7741 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     7801 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     7861 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     7921 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     7981 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     8041 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     8101 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtg
//
