LOCUS       sequence-342086-        4928 bp    DNA     circular SYN 14-APR-2025
DEFINITION  In vitro transcription of TurboID-EGFP. Parton lab clone JRS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4928)
  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 4928)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-TurboID-EGFP
FEATURES             Location/Qualifiers
     source          1..4928
                     /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(1115..1139)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(1141..1143,1144..1857)
                     /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(1141..1143,1144..1854)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1183..1204)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1444..1463)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1791..1812
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1859..1879)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     3'UTR           1904..2029
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(2116..2135)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2117..2135)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2142..2159)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2142..2158)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2151..2173)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(2300..2755)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2437..2458)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2649..2668
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2782..2886
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2887..2955,2956..3747)
                     /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(3105..3124)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3918..4506
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4407..4426
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4660..4677
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4794..4815
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4830..4835,4836..4853,4854..4860)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4868..4884
                     /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     4873..4895
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4892..4908
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4892..4908
                     /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 accatggagc agaaactgat aagcgaagag gacctgaaag acaatacagt ccccttgaag
      181 ctgattgccc ttctggcaaa tggcgaattc cacagcgggg agcaactggg cgaaacgttg
      241 ggcatgagta gagcagcgat aaacaaacat atacaaacgc tgagggattg gggtgtagac
      301 gtgttcaccg ttccgggcaa gggctatagt ttgcccgaac cgataccact tctgaacgcc
      361 aaacaaatat tgggtcaact ggatggagga tcagtagcag ttcttcctgt ggtcgactct
      421 actaaccaat atctgctgga tcggatcggg gaactgaaat ctggcgatgc atgcatagct
      481 gagtaccagc aggctggacg gggatctcga ggaaggaaat ggttttcccc gtttggggcc
      541 aacctctatc tctctatgtt ctggaggttg aaacgtggcc ctgctgccat cggcttgggg
      601 cctgtcattg gtattgttat ggcggaggcg ttgcgtaaac ttggagcgga taaagtgagg
      661 gttaaatggc caaacgattt gtatcttcaa gaccgcaagt tggctggtat actggtagaa
      721 ctcgcaggga ttacaggaga tgcggctcag attgtgatcg gtgccggtat taacgtcgcg
      781 atgcgtaggg tagaggagtc cgtggtcaat cagggttgga ttacgctgca agaggctgga
      841 atcaatctcg acagaaacac acttgcagcc acgcttattc gggaattgag ggctgctctt
      901 gaactttttg aacaggaagg gttggcacct tacctgccgc gctgggaaaa gctggataat
      961 ttcataaatc ggcccgtcaa gctcataatc ggggacaagg aaattttcgg catatcacgc
     1021 ggcatcgaca agcagggggc actgctgttg gaacaggacg gagtcataaa gccctggatg
     1081 ggtggtgaga ttagccttcg atctgcggag aaacacccag ctttcttgta caaagtggct
     1141 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     1201 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     1261 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     1321 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     1381 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     1441 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     1501 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     1561 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     1621 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     1681 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     1741 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     1801 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaacca
     1861 actttattat acatagttgc tcgagcggat ctggttacca ctaaaccagc ctcaagaaca
     1921 cccgaatgga gtctctaagc tacataatac caacttacac tttacaaaat gttgtccccc
     1981 aaaatgtagc cattcgtatc tgctcctaat aaaaaraaag tttcttcaca ttctaaaaaa
     2041 aaaaaaaaaa aaaaaaaaaa aaancccccc cccccccctg cagggccaag tcggcctcta
     2101 gaggtaccca attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa
     2161 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     2221 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     2281 agcctgaatg gcgaatggca aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     2341 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     2401 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     2461 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     2521 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     2581 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     2641 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     2701 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     2761 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     2821 aatatgtatc tgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     2881 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     2941 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     3001 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     3061 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     3121 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     3181 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     3241 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     3301 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     3361 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     3421 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     3481 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     3541 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     3601 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     3661 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     3721 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     3781 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     3841 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     3901 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     3961 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4021 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     4081 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4141 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     4201 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     4261 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     4321 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     4381 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     4441 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     4501 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     4561 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     4621 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     4681 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     4741 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     4801 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     4861 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     4921 aagctcga
//
