LOCUS       sequence-256601-        8667 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RhoB upstream msfYFP 24xMS2 RhoB downstream for CRISPR tagging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8667)
  AUTHORS   Wilson MZ, Ravindran PT, Lim WA, Toettcher JE
  TITLE     Tracing Information Flow from Erk to Target Gene Induction Reveals 
            Mechanisms of Dynamic and Combinatorial Control.
  JOURNAL   Mol Cell. 2017 Sep 7;67(5):757-769.e5. doi: 
            10.1016/j.molcel.2017.07.016. Epub 2017 Aug 17.
  PUBMED    28826673
REFERENCE   2  (bases 1 to 8667)
  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: 
            "10.1016/j.molcel.2017.07.016"; journalName: "Mol Cell"; date: 
            "2017-09-7- 7"; volume: "67"; issue: "5"; pages: "757-769.e5"
COMMENT     Sequence Label: pAAV RhoB YFP MS2
FEATURES             Location/Qualifiers
     source          1..8667
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      6175..6630
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6262..6281)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6472..6493
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6647..6666)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6766..6788
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6826..6844)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6912..7016
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7017..7085,7086..7877)
                     /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(7235..7254)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8048..8636
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8537..8556
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggccttttg ctggcctttt gctcacatgt agaggtgtca tctcctgcag gaagtggtaa
       61 ttcacgagtt aagacctggt gtgtatgaag aaactccatc ctctgtacag aaaccgtgtg
      121 cccaccggca agttccctgc gaggccgcat gcctgctttc ccagcttgaa gcttttcatc
      181 tgctccatga gagttgttgg gatagatggg tttccttgca cagtgtactt caagcagttc
      241 tctggcttta gggcgagaat cagatgtaat gtggtaggac tgggtgaagg ttgcctgccc
      301 aagctgggtg ctttcttttt cagcggagtg ccctgcacct cgctttcctt tgtcctacag
      361 ctgtgtgtct cctagattcc tccccttatc ttcttcaccc ggagacctaa gtgtgggcct
      421 tgaagagaaa gccgagaggg gactcaccca ggcccacaca atttactgtt agcctttctc
      481 agtgaaaagg ctgccctggg taagtgtcag aattcggccc agagcacacg aggggttagt
      541 tatctaaaca aagttaagaa agaacgaaac ccttggctag gtaacaaaac cgtgcaccgg
      601 tgtctgttta agtgaaaaca aaaccacgtt aaaggcgccg cccctccccc atgcttgatc
      661 acccgtagtt cgctcggaag ctagataaag ctgaagccct gggagcagcc aacgccccac
      721 aagagtgtag ctcgtctcgg aactgcccac ccagacccgg agggctccag gctgcctgct
      781 ccacgatgtc gcactcagag aggctgcgac gaaggcgggg acccggcggc cacacaaccc
      841 cacggggcgg gtcgccgcgc agactccaca gtcaacgcct ctcccagccc cgcggacagg
      901 gcagtcaatc aagctaaggc gaacccgcct tcggactgca gcgggcgacc aatcggagcc
      961 aagctccgca gcgatgagct cagccctatg gcttcccatt gggtggctat attaagaaag
     1021 tggccggact ctttaaatag cgggcgctag ggccgcagcc cgcatctgcc accgcagttc
     1081 ggttgtcgtc gtctgcgccc ctgtacccta ggtctagagt gaacccgccc cgggaaagaa
     1141 gctaggccgg gtccgcagca tcagccatca cttcgcacaa agcagcgcag ctccgggacc
     1201 gccgaggacc acagcggcgc ggcagcggcg cggcgacact cagtgcaccg tatgcccctg
     1261 cgcccctgcc gaggcaaccg gagcgccccg agagaccgcg ccgccgcggg gtccaggtgc
     1321 agttagcgag cctagcccgc agcgcgcagt cgcgggagag cggggagcgg caagcaacag
     1381 ggagcgggac ggcggcgagg cgctcgcggg cccctcctgc tgcccgcgcc cggcgagctc
     1441 atggcggcca tccgcaagaa gctggtggtg gtgggcgacg gcgcgtgcgg caagacgtgc
     1501 ctgctgatcg tgttcagtaa agacgaattc cccgaggtgt acgtgcccac cgtgttcgag
     1561 aactatgtgg cggacatcga ggtggacggc aagcaggtgg agctggcgct gtgggacacg
     1621 gcaggccagg aggactacga tcgtttacgg ccgctctcct atccggacac cgacgtcatc
     1681 cttatgtgct tctcggtgga cagcccggac tctctcgaga acatccccga gaagtgggtg
     1741 cccgaggtaa agcacttctg ccccaatgtg cccatcatcc tggtggccaa caaaaaagac
     1801 ctgcgcagcg acgagcatgt ccgcacggag ctggcccgca tgaagcagga gccagtgcgc
     1861 acggatgacg gccgcgccat ggcggtgcgc atccaagcct atgactacct cgagtgctcg
     1921 gccaagacca aggagggcgt gcgcgaggtt tttgagacgg ccacgcgcgc cgcgctgcag
     1981 aagcgctacg gatcccagaa tggctgcatc aactgctgca aggtgctagg cggatccggg
     2041 ggaagtaaag gtgaagaact gttcaccggt gttgttccga tcctggttga actggatggt
     2101 gatgttaacg gccacaaatt ctctgttcgt ggtgaaggtg aaggtgatgc aaccaacggt
     2161 aaactgaccc tgaaattcat ctgcactacc ggtaaactgc cggttccatg gccgactctg
     2221 gtgactaccc tgacctatgg tgttcagtgt ttttctcgtt acccggatca catgaagcag
     2281 catgatttct tcaaatctgc aatgccggaa ggttatgtac aggagcgcac catttctttc
     2341 aaagacgatg gcacctacaa aacccgtgca gaggttaaat ttgaaggtga tactctggtg
     2401 aaccgtattg aactgaaagg cattgatttc aaagaggacg gcaacatcct gggccacaaa
     2461 ctggaatata acttcaactc ccataacgtt tacatcaccg cagacaaaca gaagaacggt
     2521 atcaaagcta acttcaaaat tcgccataac gttgaagacg gtagcgtaca gctggcggac
     2581 cactaccagc agaacactcc gatcggtgat ggtccggttc tgctgccgga taaccactac
     2641 ctgtcctacc agtctaaact gtccaaagac ccgaacgaaa agcgcgacca catggtgctg
     2701 ctggagttcg ttactgcagc aggtatcacg cacggcatgg atgaactcta caaatgataa
     2761 gccgcgaatt cgcccttcgc gggatcctac ggtacttatt gccaagaaag cacgagcatc
     2821 agccgtgcct ccaggtcgaa tcttcaaacg acgacgatca cgcgtcgctc cagtattcca
     2881 gggttcatca gatcctacgg tacttattgc caagaaagca cgagcatcag ccgtgcctcc
     2941 aggtcgaatc ttcaaacgac gacgatcacg cgtcgctcca gtattccagg gttcatcaga
     3001 tcctacggta cttattgcca agaaagcacg agcatcagcc gtgcctccag gtcgaatctt
     3061 caaacgacga cgatcacgcg tcgctccagt attccagggt tcatcagatc ctacggtact
     3121 tattgccaag aaagcacgag catcagccgt gcctccaggt cgaatcttca aacgacgacg
     3181 atcacgcgtc gctccagtat tccagggttc atcagatcct acggtactta ttgccaagaa
     3241 agcacgagca tcagccgtgc ctccaggtcg aatcttcaaa cgacgacgat cacgcgtcgc
     3301 tccagtattc cagggttcat cagatcctac ggtacttatt gccaagaaag cacgagcatc
     3361 agccgtgcct ccaggtcgaa tcttcaaacg acgacgatca cgcgtcgctc cagtattcca
     3421 gggttcatca gatcctacgg tacttattgc caagaaagca cgagcatcag ccgtgcctcc
     3481 aggtcgaatc ttcaaacgac gacgatcacg cgtcgctcca gtattccagg gttcatcaga
     3541 tcctacggta cttattgcca agaaagcacg agcatcagcc gtgcctccag gtcgaatctt
     3601 caaacgacga cgatcacgcg tcgctccagt attccagggt tcatcagatc ctacggtact
     3661 tattgccaag aaagcacgag catcagccgt gcctccaggt cgaatcttca aacgacgacg
     3721 atcacgcgtc gctccagtat tccagggttc atcagatcct acggtactta ttgccaagaa
     3781 agcacgagca tcagccgtgc ctccaggtcg aatcttcaaa cgacgacgat cacgcgtcgc
     3841 tccagtattc cagggttcat cagatcctac ggtacttatt gccaagaaag cacgagcatc
     3901 agccgtgcct ccaggtcgaa tcttcaaacg acgacgatca cgcgtcgctc cagtattcca
     3961 gggttcatca gatcctacgg tacttattgc caagaaagca cgagcatcag ccgtgcctcc
     4021 aggtcgaatc ttcaaacgac gacgatcacg cgtcgctcca gtattccagg gttcatcaga
     4081 tctaagggcg aattcgtttg ggacgcgtcc tgcctcacgc ccttgccagc gtggctcccc
     4141 ctccttggcc cggtcgccca cgaaccggga gaaagggaga cccatgcccc caaggacacc
     4201 accagactgc ctgacatctg ctggtggctc tggctggtca cgctgagaat tagcgtgggc
     4261 accgagctcc cccttcccag tgtctgtgtg tgtccagctg tgtggcacag gcctgggcac
     4321 cctgctgagt gccaaggggt tcctgagcat ccttttctaa agagccaggc ctcaaagtgt
     4381 ggttgtgtgt gtgtacgact ccctacaccc cacccgactc ctgccccacc cccgcctctg
     4441 gtttccccag gggcacgcag agtggttgcg cccccagcag gtttgcttgt aaccagcaag
     4501 ccactactgt tgctccatgt ctgtaacata gacccctgga atggcgggag gggagggctg
     4561 gggggaggat ggggctgtta cataaataca gattttattt tcggaggcag aatggtattg
     4621 tttagtggtg tgtggggcga ccagggccca gagcagctcc tttcccaggc tgggtcaggc
     4681 gcccacccat ccaagcatga actggacttg gccatctttc cacaccctgg ggaagacatt
     4741 tgcaactgac ttggggtcga gaggaagcag ctcagacagt gtctcctggg ccaaccccca
     4801 gcgaacctcc ggcagaggat ccagggtgtg cagtggggtc acttttgcca taagcgaact
     4861 ttgtgcctgt cctacaaatg aacattgttc agctcaagag actattgtta ctgaatttat
     4921 ttaaacgctg aagctttttg ttgttgatga aagagttctt tgcacaattg tcccattgtt
     4981 tgacacccag tgtacttgtc atttgcataa gacagcattc tgaccacact tgtatgctgt
     5041 aacctcatct acttctgatg gtttttttaa aactatgatg actctaagat tacaaaaaca
     5101 actaactttt gctttgtttt cttgaaaaaa aaatgtcaac agtgtgaatt ttttctttaa
     5161 atttgtgtag catacgcagt tttggtaaag gaaggcaaca ggtattgggg tctatttaaa
     5221 ccccccctcc cacaagacaa gtctcttctg tgtgaaattt tctgtacgtt ccatgtgcag
     5281 agaaaagcct cccccctctc ccccccgcca gcccagtggt acttctaaat tgtcttgttt
     5341 tgtttttgtt ttattttttt taaataaact gacagatgac aaatggtgaa cttatgatgt
     5401 ttacataaga gttctataag ctgtgtatac agatttttat gtaaaatatt aaaagactat
     5461 gatcatgaca tttataaaca gctcctgtgg cttaatagtg tcagaagatg tctgaattgg
     5521 gggtaaggga agagttcttg tggaggtcct ttctggcagt aactgcagcc tctctatagt
     5581 agtaaatatg gccatgagcc tggcaccttt taggggctgg gtgaccctga gaggccaggt
     5641 tattcctgtc agattcccgg aggaggctaa gcttgtcgtc catcagacac ccagagtgta
     5701 ctaattattg tgaatttagt cccattaggt gaggacaaag gtcttggttt gacttcatct
     5761 ataggtcttt aagaagtgtc tgaagtttat tctgggctct actactctca cttcaacaat
     5821 aagaaaggtt gctgtttgtg aatgttaaac atagatgact ttattattat tatttaaaat
     5881 gtttttaggg gggtggttaa tcacatgtgt ggaccaggct agaatttaag agatggggaa
     5941 agcatggcag tgatggcgga tgccttcagt cccagcaaaa gcaggcaaat ctcttgagtt
     6001 ataggccagc ctggactaca gatgcagagt gggttccagg gctacacaga gaaaaaagaa
     6061 cagacaagca aaggaataga caatactccc agaaactaag ggcgcctgat gcggtatttt
     6121 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     6181 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     6241 ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     6301 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     6361 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     6421 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     6481 tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga
     6541 ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     6601 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     6661 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     6721 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     6781 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     6841 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     6901 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     6961 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     7021 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     7081 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     7141 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     7201 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     7261 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     7321 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     7381 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     7441 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     7501 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     7561 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     7621 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     7681 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     7741 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     7801 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     7861 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     7921 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     7981 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     8041 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     8101 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     8161 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     8221 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     8281 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     8341 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     8401 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     8461 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     8521 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     8581 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     8641 ccagcaacgc ggccttttta cggttcc
//
