LOCUS       sequence-342101-        4742 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro transcription of miniTurboID-EGFP. Parton lab clone JRT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4742)
  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 4742)
  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-miniTurboID-EGFP
FEATURES             Location/Qualifiers
     source          1..4742
                     /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      118..127
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             127..156
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    complement(929..953)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(955..957,958..1671)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(955..957,958..1668)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(997..1018)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1258..1277)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1605..1626
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1673..1693)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     3'UTR           1718..1843
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(1930..1949)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1931..1949)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1956..1973)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1956..1972)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1965..1987)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(2114..2569)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2251..2272)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2463..2482
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2596..2700
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2701..2769,2770..3561)
                     /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(2919..2938)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3732..4320
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4221..4240
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4474..4491
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4608..4629
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4644..4649,4650..4667,4668..4674)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4682..4698
                     /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     4687..4709
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4706..4722
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4706..4722
                     /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 accatggaac aaaagctcat ctcagaagaa gatcttattc cattgttgaa cgcgaagcaa
      181 attctcggcc aacttgatgg ggggagcgtt gcggtgctcc ccgttgtgga tagcactaat
      241 caataccttc tcgataggat tggcgagctg aaatccggcg acgcctgtat tgcagagtac
      301 cagcaggccg ggcgggggtc tcgagggcgt aagtggttct caccgttcgg tgcgaacctc
      361 tacctctcaa tgttttggcg attgaagagg gggccggcag ctatcgggtt ggggccagtg
      421 atagggattg ttatggcaga ggcgctcagg aaactcggcg cggacaaggt ccgcgtgaaa
      481 tggccaaacg atttgtacct tcaagaccgg aagttggcag gaattctggt ggagctcgcc
      541 ggaataaccg gagatgccgc acaaatagtc attggtgcag ggataaatgt tgcgatgcgt
      601 cgggtcgagg aaagcgttgt aaaccaaggg tggattacat tgcaggaggc gggcatcaat
      661 ttggatagaa atactcttgc tgcaatgttg ataagggagt tgcgcgcggc cttggaattg
      721 ttcgagcagg aaggcttggc cccgtatttg agtaggtggg agaaactgga caattttatt
      781 aaccgcccag ttaagttgat catcggcgat aaagaaattt tcggaatatc cagaggtatt
      841 gacaagcaag gcgcgctcct cctggagcag gatggggtga tcaagccgtg gatgggtggt
      901 gaaatctccc tgagaagcgc ggaaaaacac ccagctttct tgtacaaagt ggctgtgagc
      961 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     1021 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     1081 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     1141 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     1201 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     1261 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     1321 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     1381 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     1441 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     1501 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     1561 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     1621 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta accaacttta
     1681 ttatacatag ttgctcgagc ggatctggtt accactaaac cagcctcaag aacacccgaa
     1741 tggagtctct aagctacata ataccaactt acactttaca aaatgttgtc ccccaaaatg
     1801 tagccattcg tatctgctcc taataaaaar aaagtttctt cacattctaa aaaaaaaaaa
     1861 aaaaaaaaaa aaaaaaancc cccccccccc cctgcagggc caagtcggcc tctagaggta
     1921 cccaattcgc cctatagtga gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt
     1981 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     2041 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     2101 aatggcgaat ggcaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     2161 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     2221 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     2281 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     2341 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     2401 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     2461 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     2521 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     2581 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     2641 tatctgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     2701 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     2761 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     2821 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     2881 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     2941 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     3001 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     3061 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     3121 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     3181 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     3241 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     3301 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     3361 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     3421 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     3481 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     3541 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     3601 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     3661 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     3721 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3781 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     3841 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     3901 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     3961 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4021 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4081 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4141 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4201 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4261 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     4321 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     4381 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     4441 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     4501 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     4561 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     4621 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     4681 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc
     4741 ga
//
