LOCUS       sequence-435542-        5963 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GAL4-sfGFP(1-157) construct expression in mammlian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5963)
  AUTHORS   Barykina NV, Carey EM, Oliinyk OS, Nimmerjahn A, Verkhusha VV
  TITLE     Destabilized near-infrared fluorescent nanobodies enable 
            background-free targeting of GFP-based biosensors for imaging and 
            manipulation.
  JOURNAL   Nat Commun. 2024 Sep 6;15(1):7788. doi: 10.1038/s41467-024-51857-x.
  PUBMED    39242569
REFERENCE   2  (bases 1 to 5963)
  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 
            Commun."; date: "2024-09-6"; volume: "15(1):7788. doi"; pages: " 
            10.1038/s41467-024-51857-x"
COMMENT     Sequence Label: pGAL4-sfGFP(1-157)
FEATURES             Location/Qualifiers
     source          1..5963
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             914..1354
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     CDS             join(1397..1399,1400..1858)
                     /codon_start=1
                     /product="improved N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN155(I152L)
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITA"
     primer_bind     complement(1439..1460)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             1886..1915
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1931..1948
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1971..1988)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1977..2201
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2247..2675
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2334..2353)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2544..2565
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2684..2704)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2689..3018
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2869..3004
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2931..2950
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3066..3113
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3132..3506
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    3636..3757
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3673..3692)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3727..3746
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3806..3822)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3806..3822)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3819..3841)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3830..3846
                     /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)."
     promoter        complement(join(3854..3860,3861..3878,3879..3884))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3899..3920
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4037..4054)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4208..4796)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4288..4307)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4967..5758,5759..5827))
                     /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     5590..5609
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5828..5932)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttggt accatgaagc tactgtcttc tatcgaacaa gcatgcgata tttgccgact
      961 taaaaagctc aagtgctcca aagaaaaacc gaagtgcgcc aagtgtctga agaacaactg
     1021 ggagtgtcgc tactctccca aaaccaaaag gtctccgctg actagggcac atctgacaga
     1081 agtggaatca aggctagaaa gactggaaca gctatttcta ctgatttttc ctcgagaaga
     1141 ccttgacatg attttgaaaa tggattcttt acaggatata aaagcattgt taacaggatt
     1201 atttgtacaa gataatgtga ataaagatgc cgtcacagat agattggctt cagtggagac
     1261 tgatatgcct ctaacattga gacagcatag aataagtgcg acatcatcat cggaagagag
     1321 tagtaacaaa ggtcaaagac agttgactgt atcgggtgga ggatccggag gtggtggatc
     1381 aggaggaggc ggccgcgtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     1441 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgcgcggcg agggcgaggg
     1501 cgatgccacc aacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     1561 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     1621 cgaccacatg aagcgccacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     1681 gcgcaccatc agcttcaagg acgacggcac ctacaagacc cgcgccgagg tgaagttcga
     1741 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     1801 catcctgggg cacaagctgg agtacaactt caacagccac aacgtctata tcaccgccga
     1861 caagcagtaa tctagagggc ccttcgaaca aaaactcatc tcagaagagg atctgaatat
     1921 gcataccggt catcatcacc atcaccattg agtttaaacc cgctgatcag cctcgactgt
     1981 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     2041 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     2101 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     2161 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     2221 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     2281 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     2341 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     2401 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     2461 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     2521 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     2581 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     2641 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     2701 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     2761 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     2821 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     2881 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     2941 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3001 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag
     3061 cacgtgttga caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga
     3121 ggaactaaac catggccaag ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg
     3181 ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc gtggaggacg
     3241 acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc caggaccagg
     3301 tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg
     3361 agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc atgaccgaga
     3421 tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc aactgcgtgc
     3481 acttcgtggc cgaggagcag gactgacacg tgctacgaga tttcgattcc accgccgcct
     3541 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     3601 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     3661 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     3721 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     3781 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     3841 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     3901 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     3961 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4021 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4081 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4141 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4201 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4261 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4321 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4381 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4441 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4501 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4561 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4621 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4681 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4741 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4801 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4861 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4921 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     4981 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5041 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5101 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5161 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5221 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5281 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5341 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5401 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5461 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5521 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5581 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5641 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5701 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5761 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5821 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     5881 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     5941 cccgaaaagt gccacctgac gtc
//
