LOCUS       sequence-232880-        7910 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses 3xHA-tagged miniTurbo in C. elegans intestine.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7910)
  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 7910)
  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-miniTurbo_pAS32
FEATURES             Location/Qualifiers
     source          1..7910
                     /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(7263..7282)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7264..7282)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7292..7309)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7292..7308)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7301..7323)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7450..7905
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7537..7556)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7747..7768
                     /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 ctatcccact tcttaacgct aagcaaatcc ttggacaact
     5701 tgatggagga tccgtcgctg tccttccagt tgttgattcc accaaccaat accttcttga
     5761 tcgtatcgga gagcttaagt ctggagacgc ttgcatcgct gaataccagc aggctggtag
     5821 aggtaagttt aaacagttcg gtactaacta accatacata tttaaatttt caggaagtcg
     5881 tggccgtaaa tggtttagtc ctttcggcgc caacctctac cttagtatgt tttggcgtct
     5941 gaaaagagga cctgcggcga taggcttggg tccagtgatc ggtattgtta tggccgaggc
     6001 gctgcgaaag ctgggagctg ataaggttcg tgttaagtgg ccaaacgatc tttaccttca
     6061 agaccgtaag cttgctggaa tccttgtcga gcttgctgga atcaccggag acgctgctca
     6121 aatcgttatc ggagctggaa tcaacgttgc tatgcgtcgt gttgaggtaa gtttaaacat
     6181 gattttacta actaactaat ctgatttaaa ttttcaggag tctgttgtta accaaggatg
     6241 gatcactctt caagaggctg gaatcaacct tgatcgtaac acccttgctg ccatgcttat
     6301 ccgtgagctt cgtgctgccc ttgaactttt cgagcaagag ggacttgctc catacctttc
     6361 tcgttgggag aagcttgata acttcatcaa ccgcccagtt aagcttatca tcggagataa
     6421 ggaaatcttc ggaatctctc gcggaatcga taagcaagga gctcttcttc ttgagcaaga
     6481 tggagttatt aaaccatgga tgggaggaga aatttccctt cgttccgccg agaagtaacc
     6541 cgggcatctc gcgcccgtgc ctctgacttc taagtccaat tactcttcaa catccctaca
     6601 tgctctttct ccctgtgctc ccacccccta tttttgttat tatcaaaaaa cttctcttaa
     6661 tttctttgtt ttttagcttc ttttaagtca cctctaacaa tgaaattgtg tagattcaaa
     6721 aatagaatta attcgtaata aaaagtcgaa aaaaattgtg ctccctcccc ccattaataa
     6781 taattctatc ccaaaatcta cacaatgttc tgtgtacact tcttatgttt tttacttctg
     6841 ataaattttt ttgaaacatc atagaaaaaa ccgcacacaa aataccttat catatgttac
     6901 gtttcagttt atgaccgcaa tttttatttc ttcgcacgtc tgggcctctc atgacgtcaa
     6961 atcatgctca tcgtgaaaaa gttttggagt atttttggaa tttttcaatc aagtgaaagt
     7021 ttatgaaatt aattttcctg cttttgcttt ttggggtttc ccctattgtt tgtcaagatt
     7081 tcgaggacgg cgtttttctt gctaaaatca caagtattga tgagcacgat gcaagaaaga
     7141 tcggaagaag gtttgggttt gaggctcagt ggaaggtgag tagaagttga taatttgaaa
     7201 gtggagtagt gtctatgggg tttttgcctt aaatctcgag ggggggcccg gtacccaatt
     7261 cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact
     7321 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     7381 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     7441 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     7501 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     7561 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     7621 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     7681 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     7741 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     7801 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     7861 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
//
