LOCUS       sequence-389480-        5623 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses RHOB GTPase with tripartite split GFP10 tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5623)
  AUTHORS   Koraichi F, Gence R, Bouchenot C, Grosjean S, Lajoie-Mazenc I, Favre
            G, Cabantous S
  TITLE     High-content tripartite split-GFP cell-based assays to screen for 
            modulators of small GTPase activation.
  JOURNAL   J Cell Sci. 2018 Jan 8;131(1). pii: jcs.210419. doi: 
            10.1242/jcs.210419.
  PUBMED    29192060
REFERENCE   2  (bases 1 to 5623)
  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: "J Cell 
            Sci."; date: "2018-01-8"; volume: "131(1). pii: jcs.210419. doi"; 
            pages: " 10.1242/jcs.210419"
COMMENT     Sequence Label: pcDNA GFP10-RHOB
FEATURES             Location/Qualifiers
     source          1..5623
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(65..84)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        256..635
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        636..839
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     790..810
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     884..903
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        884..902
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      922..931
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1652..1669)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1658..1882
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1928..2356
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2015..2034)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2225..2246
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2365..2385)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2370..2699
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2550..2685
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2612..2631
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2747..2794
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2813..3187
                     /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    3317..3438
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3354..3373)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3408..3427
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3487..3503)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3487..3503)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3500..3522)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3511..3527
                     /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(3535..3541,3542..3559,3560..3565))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3580..3601
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3718..3735)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3889..4477)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3969..3988)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4648..5439,5440..5508))
                     /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     5271..5290
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5509..5613)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc
       61 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
      121 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
      181 acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg
      241 ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca attacggggt
      301 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
      361 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
      421 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
      481 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
      541 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
      601 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
      661 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
      721 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
      781 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
      841 tctggctaac tagagaaccc actgcttact ggcttatcga aattaatacg actcactata
      901 gggagaccca agctggctag cgccaccatg ggcgacctgc ccgacgacca ctacctgtcc
      961 acccagacca tcctgagcaa ggacctgaac atcgatggtg gcggtggctc tggaggtggt
     1021 gggtcctccg gagcggccat ccgcaagaag ctggtggtgg tgggcgacgg cgcgtgtggc
     1081 aagacgtgcc tgctgatcgt gttcagtaag gacgagttcc ccgaggtgta cgtgcccacc
     1141 gtcttcgaga actatgtggc cgacattgag gtggacggca agcaggtgga gctggcgctg
     1201 tgggacacgg cgggccagga ggactacgac cgcctgcggc cgctctccta cccggacacc
     1261 gacgtcattc tcatgtgctt ctcggtggac agcccggact cgctggagaa catccccgag
     1321 aagtgggtcc ccgaggtgaa gcacttctgt cccaatgtgc ccatcatcct ggtggccaac
     1381 aaaaaagacc tgcgcagcga cgagcatgtc cgcacagagc tggcccgcat gaagcaggaa
     1441 cccgtgcgca cggatgacgg ccgcgccatg gccgtgcgca tccaagccta cgactacctc
     1501 gagtgctctg ccaagaccaa ggaaggcgtg cgcgaggtct tcgagacggc cacgcgcgcc
     1561 gcgctgcaga agcgctacgg ctcccagaac ggctgcatca actgctgcaa ggtgctataa
     1621 tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     1681 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     1741 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     1801 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     1861 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     1921 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     1981 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     2041 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     2101 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     2161 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     2221 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     2281 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     2341 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     2401 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     2461 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2521 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     2581 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     2641 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     2701 agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgttg acaattaatc
     2761 atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa ccatggccaa
     2821 gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg
     2881 gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg gtgtggtccg
     2941 ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg acaacaccct
     3001 ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc
     3061 cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc agccgtgggg
     3121 gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg ccgaggagca
     3181 ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     3241 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     3301 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     3361 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     3421 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     3481 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     3541 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     3601 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     3661 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     3721 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     3781 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     3841 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     3901 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     3961 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4021 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4081 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4141 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4201 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4261 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4321 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4381 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4441 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     4501 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     4561 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     4621 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     4681 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     4741 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     4801 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     4861 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     4921 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     4981 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     5041 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     5101 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     5161 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     5221 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     5281 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     5341 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     5401 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     5461 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     5521 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     5581 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccc
//
