LOCUS Exported 7459 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pBabe-Puro-RhoA FLARE.sc Biosensor SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7459) AUTHORS Pertz O, Hodgson L, Klemke RL, Hahn KM TITLE Spatiotemporal dynamics of RhoA activity in migrating cells. JOURNAL Nature. 2006 Apr 20. 440(7087):1069-72. PUBMED 16547516 REFERENCE 2 (bases 1 to 7459) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7459 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 7..24 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 289..1005 /codon_start=1 /product="cyan variant of GFP" /label=CFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 1078..1794 /codon_start=1 /product="enhanced variant of YFP (Heikal et al., 2001)" /label=Citrine /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 1801..2382 /codon_start=1 /product="human Rho family small GTPase RhoA" /label=RhoA /translation="MAAIRKKLVIVGDGACGKTCLLIVFSKDQFPEVYVPTVFENYVAD IEVDGKQVELALWDTAGQEDYDRLRPLSYPDTDVILMCFSIDSPDSLENIPEKWTPEVK HFCPNVPIILVGNKKDLRNDEHTRRELAKMKQEPVKPEEGRDMANRIGAFGYMECSAKT KDGVREVFEMATRAALQARRGKKKSGCLVL" primer_bind complement(2397..2417) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2402..2731 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2582..2717 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2644..2663 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2741..3340 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(2741..2760) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 3237..3257 /label=Puro-F /note="Puromycin resistance gene, forward primer" LTR 3561..4030 /note="long terminal repeat from Moloney murine leukemia virus" primer_bind complement(4160..4177) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4331..4919) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4411..4430) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5079..5939) /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 5702..5721 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5940..6044) /gene="bla" /label=AmpR promoter LTR 6075..6544 /note="long terminal repeat from Moloney murine leukemia virus" misc_feature 6609..6808 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" CDS 7009..7425 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 7355..7377 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 7393..7409 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" ORIGIN 1 atggcacacc atcaccacca tcacggtagt ggcatcctgg aggacctcaa tatgctctac 61 atccggcaga tggcactcag cctggaggac acagagctgc agaggaaact agatcatgag 121 atccggatga gggatggggc ctgcaagctg ctggcagcct gctcccagcg agagcaggct 181 ctggaagcca ccaagagcct gctggtgtgc aacagccgta ttctcagcta catgggtgag 241 ctgcagcggc gaaaggaggc ccaggtgctg gagaagacag gcgggggaat ggtgagcaag 301 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac 361 ggccacaggt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc 421 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc 481 ctgacctggg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc 541 ttcaagtccg ccatgcccga aggctacgtc caggagcgta ccatcttctt caaggacgac 601 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc 661 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac 721 aactacatca gccacaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc 781 cacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag 841 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc 901 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc 961 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagggatc tacttctggt 1021 tctggtaaac ctggttctgg tgaaggttct actaaaggtg gtagtggcgc ggccgctatg 1081 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc 1141 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc 1201 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc 1261 gtgaccacct tcggctacgg cctgatgtgc ttcgcccgct accccgacca catgaagcag 1321 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc 1381 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg 1441 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag 1501 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc 1561 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac 1621 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac 1681 ctgagctacc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg 1741 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagggggga 1801 atggctgcca tccggaagaa actggtgatt gttggtgatg gagcctgtgg aaagacatgc 1861 ttgctcatag tcttcagcaa ggaccagttc ccagaggtgt atgtgcccac agtgtttgag 1921 aactatgtgg cagatatcga ggtggatgga aagcaggtag agttggcttt gtgggacaca 1981 gctgggcagg aagattatga tcgcctgagg cccctctcct acccagatac cgatgttata 2041 ctgatgtgtt tttccatcga cagccctgat agtttagaaa acatcccaga aaagtggacc 2101 ccagaagtca agcatttctg tcccaacgtg cccatcatcc tggttgggaa taagaaggat 2161 cttcggaatg atgagcacac aaggcgggag ctagccaaga tgaagcagga gccggtgaaa 2221 cctgaagaag gcagagatat ggcaaacagg attggcgctt ttgggtacat ggagtgttca 2281 gcaaagacca aagatggagt gagagaggtt tttgaaatgg ctacgagagc tgctctgcaa 2341 gctagacgtg ggaagaaaaa atctggttgc cttgtcttgt gataactcga ccctgtggaa 2401 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag 2461 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag 2521 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc 2581 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt 2641 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg 2701 aggctttttt ggaggcctag gcttttgcaa aaagcttacc atgaccgagt acaagcccac 2761 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt 2821 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac 2881 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc 2941 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt 3001 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca 3061 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac 3121 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg 3181 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa 3241 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg 3301 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga cgcccgcccc acgacccgca 3361 gcgcccgacc gaaaggagcg cacgacccca tgcatcgata aaataaaaga ttttatttag 3421 tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc tagcttaagt 3481 aacgccattt tgcaaggcat ggaaaaatac ataactgaga atagagaagt tcagatcaag 3541 gtcaggaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc 3601 ctgccccggc tcagggccaa gaacagatgg aacagctgaa tatgggccaa acaggatatc 3661 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc 3721 cagccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa 3781 atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc 3841 ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggggcg ccagtcctcc 3901 gattgactga gtcgcccggg tacccgtgta tccaataaac cctcttgcag ttgcatccga 3961 cttgtggtct cgctgttcct tgggagggtc tcctctgagt gattgactac ccgtcagcgg 4021 gggtctttca catgcagcat gtatcaaaat taatttggtt ttttttctta agtatttaca 4081 ttaaatggcc atagttgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta 4141 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc 4201 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg 4261 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt 4321 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa 4381 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct 4441 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc 4501 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg 4561 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct 4621 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag 4681 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga 4741 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga 4801 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg 4861 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 4921 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 4981 ggattttggt catgagatta tcaaaaagga tcttcaccta gatccttttg cggccgcaaa 5041 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag 5101 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg 5161 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga 5221 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag 5281 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa 5341 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc 5401 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca 5461 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg 5521 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat 5581 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc 5641 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg 5701 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg 5761 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt 5821 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca 5881 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata 5941 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac 6001 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa 6061 gtgccacctg cagcctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc 6121 cggctcaggg ccaagaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt 6181 aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc ggtccagccc 6241 tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc 6301 ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc 6361 tccccgagct caataaaaga gcccacaacc cctcactcgg ggcgccagtc ctccgattga 6421 ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg 6481 gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct 6541 ttcatttggg ggctcgtccg ggatcgggag acccctgccc agggaccacc gacccaccac 6601 cgggaggcaa gctggccagc aacttatctg tgtctgtccg attgtctagt gtctatgact 6661 gattttatgc gcctgcgtcg gtactagtta gctaactagc tctgtatctg gcggacccgt 6721 ggtggaactg acgagttctg aacacccggc cgcaaccctg ggagacgtcc cagggacttt 6781 gggggccgtt tttgtggccc gacctgagga agggagtcga tgtggaatcc gaccccgtca 6841 ggatatgtgg ttctggtagg agacgagaac ctaaaacagt tcccgcctcc gtctgaattt 6901 ttgctttcgg tttggaaccg aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt 6961 ctgtgttgtc tctgtctgac tgtgtttctg tatttgtctg aaaattaggg ccagactgtt 7021 accactccct taagtttgac cttaggtcac tggaaagatg tcgagcggat cgctcacaac 7081 cagtcggtag atgtcaagaa gagacgttgg gttaccttct gctctgcaga atggccaacc 7141 tttaacgtcg gatggccgcg agacggcacc tttaaccgag acctcatcac ccaggttaag 7201 atcaaggtct tttcacctgg cccgcatgga cacccagacc aggtccccta catcgtgacc 7261 tgggaagcct tggcttttga cccccctccc tgggtcaagc cctttgtaca ccctaagcct 7321 ccgcctcctc ttcctccatc cgccccgtct ctcccccttg aacctcctcg ttcgaccccg 7381 cctcgatcct ccctttatcc agccctcact ccttctctag gcgccggccg gatcccagtg 7441 tggtggtacg taggaattc //