LOCUS       sequence-151512-        7235 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Membrane localized GFP-tagged LOVpep.  Expresses in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7235)
  AUTHORS   Wagner E, Glotzer M
  TITLE     Local RhoA activation induces cytokinetic furrows independent of 
            spindle position and cell cycle stage.
  JOURNAL   J Cell Biol. 2016 Jun 20;213(6):641-9. doi: 10.1083/jcb.201603025. 
            Epub 2016 Jun 13.
  PUBMED    27298323
REFERENCE   2  (bases 1 to 7235)
  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"; doi: 
            "10.1083/jcb.201603025"; journalName: "J Cell Biol"; date: 
            "2016-06-20- 20"; volume: "213"; issue: "6"; pages: "641-9"
COMMENT     Sequence Label: Stargazin-GFP-LOVpep
FEATURES             Location/Qualifiers
     source          1..7235
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(43..62)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        234..613
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        614..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    819..837
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    840..858
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     868..892
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1997..2710
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2027..2055)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2650..2670
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(3246..3263)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3252..3476
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3522..3950
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3609..3628)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3819..3840
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3959..3979)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3964..4293
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4144..4279
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4206..4225
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4341..4388
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4407..4781
                     /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    4911..5032
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4948..4967)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5002..5021
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5081..5097)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5081..5097)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5094..5116)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5105..5121
                     /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(5129..5135,5136..5153,5154..5159))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5174..5195
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5312..5329)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5483..6068)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5563..5582)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6239..7030,7031..7099))
                     /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     6862..6881
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7100..7204)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
       61 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
      121 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
      181 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg
      241 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
      301 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      361 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      421 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      481 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      541 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      601 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      661 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggaacca
      721 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
      781 taggcgtgta cggtgggagg tctatataag cagagctctc cctatcagtg atagagatct
      841 ccctatcagt gatagagatc gtcgacgagc tcgtttagtg aaccgtcaga tcgcctggag
      901 acgccatcca cgctgttttg acctccatag aagacaccgg gaccgatcca gcctccggac
      961 tctagcgttt aaacttaaga tggggctgtt tgatcgaggt gttcaaatgc ttttaaccac
     1021 cgttggtgct ttcgctgcct tcagcttgat gaccatcgct gtgggaaccg actattggct
     1081 gtactccaga ggggtttgca agaccaaaag tgtcagtgag aatgaaacca gcaagaagaa
     1141 cgaggaagtt atgacccatt ccggattatg gagaacctgc tgcctcgaag ggaacttcaa
     1201 aggtctgtgc aagcaaatcg accactttcc ggaagacgcg gactacgaag ctgacaccgc
     1261 agagtatttc ctccgggccg tgagggcctc gagtatcttc ccgatcctga gtgtgatcct
     1321 gcttttcatg ggtggcctct gcatcgcagc cagcgagttc tacaaaccgg tgaactgcag
     1381 gaagagctcg aagaattcga aaagcttgac acgccacaac atcatcctga gtgctggcat
     1441 cttcttcgtg tctgcaggtc ttagtaatat catcgggatc atcgtgtata tatcagccaa
     1501 tgctggagac ccctccaaga gtgactccaa aaagaacagc tactcctacg gctggtcctt
     1561 ctacttcggg gccctgtcct tcatcatcgc cgagatggtc ggggtgctgg ccgtgcacat
     1621 gtttatcgac cgccacaaac agctgcgggc cacggcccgc gccaccgact acctccaggc
     1681 ctccgccatc acccgcatcc ccagctaccg ctaccgctac cagcgccgca gccgctccag
     1741 ctcgcgctcc accgagccct ctcactccag agacgcctcg cccgtgggcg tgaagggctt
     1801 caacaccctg ccgtccacgg agatctccat gtacaccctc agtagggacc ccctgaaggc
     1861 tgccaccacg cccaccgcca cctacaactc ggacagggat aacagctttc tccaggtcca
     1921 caactgtatc cagaaggaca gcaaggactc tttccacgcc aacacagcca accgccggac
     1981 cacgcccgta ggatccatga gtaaaggaga agaacttttc actggagttg tcccaattct
     2041 tgttgaatta gatggtgatg ttaatgggca caaattttct gtcagtggag agggtgaagg
     2101 tgatgcaaca tacggaaaac ttacccttaa atttatttgc actactggaa aactacctgt
     2161 tccatggcca acacttgtca ctactttctc ttatggtgtt caatgctttt caagataccc
     2221 agatcatatg aaacggcatg actttttcaa gagtgccatg cccgaaggtt atgtacagga
     2281 aagaactata tttttcaaag atgacgggaa ctacaagaca cgtgctgaag tcaagtttga
     2341 aggtgatacc cttgttaata gaatcgagtt aaaaggtatt gattttaaag aagatggaaa
     2401 cattcttgga cacaaattgg aatacaacta taactcacac aatgtataca tcatggcaga
     2461 caaacaaaag aatggaatca aagttaactt caaaattaga cacaacattg aagatggaag
     2521 cgttcaacta gcagaccatt atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt
     2581 accagacaac cattacctgt ccacacaatc tgccctttcg aaagatccca acgaaaagag
     2641 agaccacatg gtccttcttg agtttgtaac agctgctggg attacacatg gcatggacga
     2701 gctgtacaag tctagaggtg gatctggagg ttcaggtgga agtcctaggt tggctgctgc
     2761 acttgaacgt attgagaaga actttgtcat tactgaccca aggttgccag ataatcccat
     2821 tatattcgcg tccgatagtt tcttgcagtt gacagaatat agccgtgaag aaattttggg
     2881 aagaaactgc aggtttctac aaggtcctga aactgatcgc gcgacagtga gaaaaattag
     2941 agatgccata gataaccaaa cagaggtcac tgttcagctg attaattata caaagagtgg
     3001 taaaaagttc tggaacctct ttcacttgca gcctatgcga gatcagaagg gagatgtcca
     3061 gtactttatt ggggttcagt tggatggaac tgagcatgtc cgagatgctg ccgagagaga
     3121 ggcagtcatg ctggcgaaga aaactgcaga agagattgat aaagcggtgg atacctgggt
     3181 gtaagcttct cgagtcatgt aattagttat gtcacctaga gggcccgttt aaacccgctg
     3241 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     3301 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     3361 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     3421 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     3481 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     3541 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3601 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3661 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3721 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3781 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3841 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     3901 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     3961 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4021 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4081 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4141 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4201 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     4261 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     4321 ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat
     4381 aatacgacaa ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt ccggtgctca
     4441 ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg ttctcccggg
     4501 acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg
     4561 cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg
     4621 acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac gcctccgggc
     4681 cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg cgcgacccgg
     4741 ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtgcta cgagatttcg
     4801 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     4861 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     4921 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     4981 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5041 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     5101 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     5161 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     5221 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5281 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5341 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5401 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5461 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5521 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5581 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5641 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     5701 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5761 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5821 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5881 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     5941 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6001 aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6061 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6121 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6181 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6241 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6301 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6361 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6421 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6481 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6541 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     6601 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     6661 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     6721 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     6781 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     6841 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6901 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     6961 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7021 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7081 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7141 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7201 cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
