LOCUS       sequence-342268-        4244 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro transcription of nls-TurboID. Parton lab clone JST.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4244)
  AUTHORS   Xiong Z, Lo HP, McMahon KA, Martel N, Jones A, Hill MM, Parton RG, 
            Hall TE
  TITLE     In vivo proteomic mapping through GFP-directed proximity-dependent 
            biotin labelling in zebrafish.
  JOURNAL   Elife. 2021 Feb 16;10. pii: 64631. doi: 10.7554/eLife.64631.
  PUBMED    33591275
REFERENCE   2  (bases 1 to 4244)
  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: "2021-02-16"; volume: "10. pii: 64631. doi"; pages: " 
            10.7554/eLife.64631"
COMMENT     Sequence Label: pT3TS-NLS-TurboID
FEATURES             Location/Qualifiers
     source          1..4244
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     5'UTR           44..86
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     protein_bind    93..117
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      121..130
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             133..153
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(158..182)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1141..1170
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    complement(1174..1194)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     3'UTR           1219..1344
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(1432..1451)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1433..1451)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1458..1475)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1458..1474)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1467..1489)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(1616..2071)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1753..1774)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1965..1984
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2098..2202
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2203..2271,2272..3063)
                     /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(2421..2440)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3234..3822
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3723..3742
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3976..3993
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4110..4131
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4146..4151,4152..4169,4170..4176)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4184..4200
                     /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     4189..4211
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4208..4224
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4208..4224
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aattaaccct cactaaaggg aacaaaagct ggagctaagc ttgcttgttc tttttgcaga
       61 agctcagaat aaacgctcaa ctttggcaga tctcaagttt gtacaaaaaa gcaggctgcc
      121 gccaccatgg ctccaaagaa gaagcgtaag gtacggtacc cagctttctt gtacaaagtg
      181 gtgaaagaca atacagtccc cttgaagctg attgcccttc tggcaaatgg cgaattccac
      241 agcggggagc aactgggcga aacgttgggc atgagtagag cagcgataaa caaacatata
      301 caaacgctga gggattgggg tgtagacgtg ttcaccgttc cgggcaaggg ctatagtttg
      361 cccgaaccga taccacttct gaacgccaaa caaatattgg gtcaactgga tggaggatca
      421 gtagcagttc ttcctgtggt cgactctact aaccaatatc tgctggatcg gatcggggaa
      481 ctgaaatctg gcgatgcatg catagctgag taccagcagg ctggacgggg atctcgagga
      541 aggaaatggt tttccccgtt tggggccaac ctctatctct ctatgttctg gaggttgaaa
      601 cgtggccctg ctgccatcgg cttggggcct gtcattggta ttgttatggc ggaggcgttg
      661 cgtaaacttg gagcggataa agtgagggtt aaatggccaa acgatttgta tcttcaagac
      721 cgcaagttgg ctggtatact ggtagaactc gcagggatta caggagatgc ggctcagatt
      781 gtgatcggtg ccggtattaa cgtcgcgatg cgtagggtag aggagtccgt ggtcaatcag
      841 ggttggatta cgctgcaaga ggctggaatc aatctcgaca gaaacacact tgcagccacg
      901 cttattcggg aattgagggc tgctcttgaa ctttttgaac aggaagggtt ggcaccttac
      961 ctgccgcgct gggaaaagct ggataatttc ataaatcggc ccgtcaagct cataatcggg
     1021 gacaaggaaa ttttcggcat atcacgcggc atcgacaagc agggggcact gctgttggaa
     1081 caggacggag tcataaagcc ctggatgggt ggtgagatta gccttcgatc tgcggagaaa
     1141 gagcagaaac tgataagcga agaggacctg taacaacttt attatacata gttgctcgag
     1201 cggatctggt taccactaaa ccagcctcaa gaacacccga atggagtctc taagctacat
     1261 aataccaact tacactttac aaaatgttgt cccccaaaat gtagccattc gtatctgctc
     1321 ctaataaaaa raaagtttct tcacattcta aaaaaaaaaa aaaaaaaaaa aaaaaaaann
     1381 cccccccccc cccctgcagg gccaagtcgg cctctagagg tacccaattc gccctatagt
     1441 gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     1501 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     1561 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcaaatt
     1621 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     1681 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     1741 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     1801 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     1861 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     1921 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     1981 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     2041 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     2101 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatctgct catgagacaa
     2161 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     2221 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     2281 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     2341 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     2401 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     2461 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     2521 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     2581 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     2641 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     2701 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     2761 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     2821 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     2881 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     2941 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     3001 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     3061 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     3121 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     3181 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     3241 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3301 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     3361 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     3421 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3481 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     3541 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     3601 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3661 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3721 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3781 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3841 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3901 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     3961 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     4021 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     4081 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     4141 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     4201 tttcacacag gaaacagcta tgaccatgat tacgccaagc tcga
//
