LOCUS       sequence-342067-        4751 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro transcription of BASU-EGFP. Parton lab clone JRR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4751)
  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 4751)
  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-BASU-EGFP
FEATURES             Location/Qualifiers
     source          1..4751
                     /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(938..962)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(964..966,967..1680)
                     /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(964..966,967..1677)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1006..1027)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1267..1286)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1614..1635
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1682..1702)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     3'UTR           1727..1852
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(1939..1958)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1940..1958)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1965..1982)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1965..1981)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1974..1996)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(2123..2578)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2260..2281)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2472..2491
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2605..2709
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2710..2778,2779..3570)
                     /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(2928..2947)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3741..4329
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4230..4249
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4483..4500
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4617..4638
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4653..4658,4659..4676,4677..4683)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4691..4707
                     /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     4696..4718
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4715..4731
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4715..4731
                     /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 aaaagttgat aagcgaagag gatctcggta agctcagtga aagcgaaatc
      181 aggtttgggt tgaaaacgga agtaatgggc cagcacctta tttaccatga tgtcctctca
      241 tccacccaaa agactgctca cgaattggct aacaacaacg ctcctgaagg aactctggtc
      301 gtcgcggaca agcagactgc gggaagaggt gggatgtcaa gagtttggca ctcacaggaa
      361 gggaatgggg tatggatgtc acttattctt cgtccagaca taccacttca gaagacacct
      421 cagttgaccc tcctggcggc cgtcgcagtc gtccagggaa tcgaagaggc tgccggaatc
      481 caaacagata ttaagtggcc aaatgatata ttgataaatg gcaaaaaaac tgtcggtatt
      541 cttaccgaaa tgcaggccga ggaagacaga gtccgtagcg taatcatagg catcggcatt
      601 aatgttaatc aacaaccaaa tgattttccc gatgaactta aagatattgc cactagcttg
      661 tcccaggcag caggcgagaa aatagaccgt gcaggcgtga tccaacatat actcctttgc
      721 tttgagaagc gttaccgaga ttacatgact catggtttta ctccgatcaa gttgctgtgg
      781 gagagttatg ctctgggcat tggcacgaat atgcgggctc gcacactcaa cgggactttt
      841 tacggtaagg ccttggggat tgatgacgaa ggggtactcc tgctcgaaac caacgaaggg
      901 attaaaaaga tttacagtgc agatatttca ctgaggcacc cagctttctt gtacaaagtg
      961 gctgtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     1021 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     1081 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     1141 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     1201 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     1261 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     1321 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     1381 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     1441 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     1501 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     1561 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     1621 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     1681 ccaactttat tatacatagt tgctcgagcg gatctggtta ccactaaacc agcctcaaga
     1741 acacccgaat ggagtctcta agctacataa taccaactta cactttacaa aatgttgtcc
     1801 cccaaaatgt agccattcgt atctgctcct aataaaaaga aagtttcttc acattctaaa
     1861 aaaaaaaaaa aaaaaaaaaa aaaaaanccc cccccccccc ctgcagggcc aagtcggcct
     1921 ctagaggtac ccaattcgcc ctatagtgag tcgtattaca attcactggc cgtcgtttta
     1981 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     2041 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     2101 cgcagcctga atggcgaatg gcaaattgta agcgttaata ttttgttaaa attcgcgtta
     2161 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     2221 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     2281 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     2341 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     2401 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     2461 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     2521 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     2581 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     2641 tcaaatatgt atctgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     2701 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     2761 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     2821 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     2881 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     2941 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     3001 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     3061 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     3121 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     3181 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     3241 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     3301 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     3361 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     3421 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     3481 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     3541 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     3601 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     3661 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     3721 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     3781 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     3841 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     3901 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     3961 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     4021 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     4081 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     4141 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4201 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4261 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     4321 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     4381 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     4441 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     4501 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     4561 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     4621 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     4681 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     4741 gccaagctcg a
//
