LOCUS Exported 7011 bp ds-DNA circular SYN 11-MAR-2022 DEFINITION Retroviral expression of Cdc42 splice variant 1. ACCESSION . VERSION . KEYWORDS MIGR1-Cdc42-1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7011) TITLE Retroviral expression vectors for Cdc42 splice variants JOURNAL Unpublished REFERENCE 2 (bases 1 to 7011) AUTHORS . TITLE Direct Submission JOURNAL Exported Mar 11, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7011 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 147..166 /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 406..427 /label=EGFP-N /note="EGFP, reverse primer" misc_feature 484..1036 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(646..665) /label=IRES-F /note="IRES internal ribosome entry site, forward primer" primer_bind 875..892 /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" CDS complement(1058..1633) /codon_start=1 /product="human Rho family small GTPase Cdc42" /label=Cdc42 /translation="MQTIKCVVVGDGAVGKTCLLISYTTNKFPSEYVPTVFDNYAVTVM IGGEPYTLGLFDTAGQEDYDRLRPLSYPQTDVFLVCFSVVSPSSFENVKEKWVPEITHH CPKTPFLLVGTQIDLRDDPSTIEKLAKNKQKPITPETAEKLARDLKAVKYVECSALTQK GLKNVFDEAILAALEPPEPKKSRRCVLL" CDS complement(1652..2068) /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind complement(1668..1684) /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" primer_bind complement(1700..1722) /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" misc_feature 2135..2476 /label=MESV Psi /note="packaging signal of murine embryonic stem cell virus" LTR 2540..3056 /label=5' LTR /note="5' long terminal repeat from murine embryonic stem cell virus" primer_bind complement(3256..3275) /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind complement(3403..3425) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(3619..3638) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3738..3760 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(3798..3816) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3884..3988 /gene="bla" /label=AmpR promoter CDS 3989..4849 /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(4207..4226) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5020..5608 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5509..5528 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5762..5779 /label=L4440 /note="L4440 vector, forward primer" protein_bind 5896..5917 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5932..5962 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5970..5986 /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)." LTR 6148..6662 /label=3' LTR /note="3' long terminal repeat from murine embryonic stem cell virus" CDS complement(6764..472) /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(6809..6830) /label=EGFP-C /note="EGFP, forward primer" ORIGIN 1 gcttgtcggc catgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca 61 ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt 121 cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg 181 tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt 241 ggtcggggta gcggctgaag cactgcacgc cgtaggtcag ggtggtcacg agggtgggcc 301 agggcacggg cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg 361 catcgccctc gccctcgcca gacacgctga acttgtggcc gtttacgtcg ccgtccagct 421 cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc atggttgtgg 481 ccatattatc atcgtgtttt tcaaaggaaa accacgtccc cgtggttcgg ggggcctaga 541 cgttttttta acctcgacta aacacatgta aagcatgtgc accgaggccc cagatcagat 601 cccatacaat ggggtacctt ctgggcatcc ttcagcccct tgttgaatac gcttgaggag 661 agccatttga ctctttccac aactatccaa ctcacaacgt ggcactgggg ttgtgccgcc 721 tttgcaggtg tatcttatac acgtggcttt tggccgcaga ggcacctgtc gccaggtggg 781 gggttccgct gcctgcaaag ggtcgctaca gacgttgttt gtcttcaaga agcttccaga 841 ggaactgctt ccttcacgac attcaacaga ccttgcattc ctttggcgag aggggaaaga 901 cccctaggaa tgctcgtcaa gaagacaggg ccaggtttcc gggccctcac attgccaaaa 961 gacggcaata tggtggaaaa taacatatag acaaacgcac accggcctta ttccaagcgg 1021 cttcggccag taacgttagg gggggggggc ggaattctca tagcagcaca cacctgcggc 1081 tcttcttcgg ttctggaggc tccagggcag ccaatattgc ttcgtcaaat acattcttta 1141 ggcctttctg tgtaagtgca gaacactcca catacttgac agccttcagg tcacgggcca 1201 gcttttcagc agtctctgga gtgataggct tctgtttgtt cttggcaagt ttctcaatag 1261 tagaggggtc atctctgaga tcaatttgag tcccaacaag caagaaagga gtctttggac 1321 agtggtgagt tatctcaggc acccactttt ctttcacgtt ttcaaatgaa gatggagaga 1381 ccactgaaaa acagactaga aatacatctg tttgtggata actcagcggt cgtaatctgt 1441 cataatcctc ttgccctgca gtatcaaaaa gtccaagagt atatggttct ccaccaatca 1501 taactgtgac tgcatagttg tcaaaaacag tcggtacata ttccgatgga aatttgtttg 1561 ttgtgtagga tatcaggaga catgttttac caacagcacc atcgcccaca acaacacact 1621 taattgtctg catggtggca gatctaattc cggcgcctag agaaggagtg agggctggat 1681 aaagggagga tcgaggcggg gtcgaacgag gaggttcaag ggggagagac ggggcggatg 1741 gaggaagagg aggcggaggc ttagggtgta caaagggctt gacccaggga ggggggtcaa 1801 aagccaaggc ttcccaggtc acgatgtagg ggacctggtc tgggtgtcca tgcgggccag 1861 gtgaaaagac cttgatctta acctgggtga tgaggtctcg gttaaaggtg ccgtctcgcg 1921 gccatccgac gttaaaggtt ggccattctg cagagcagaa ggtaacccaa cgtctcttct 1981 tgacatctac cgactggttg tgagcgatcc gctcgacatc tttccagtga cctaaggtca 2041 aacttaaggg agtggtaaca gtctggccct aattttcaga caaatacaga aacacagtca 2101 gacagagaca acacagaacg atgctgcagc gctgcagcag acaagacgcg cggcttcggt 2161 tccaaaccga aagcaaaaat tcagacggag gcgggaactg ttttaggttc tcgtctccta 2221 ccagaaccac atatcctgac ggggtcggat tccacatcga ctcccttcct caggtcgggc 2281 cacaaaaacg gcccccaaag tccctgggac gtctcccagg gttgcggccg ggtgttcaga 2341 actcgtcagt tccaccacgg gtccgccaga tacagagcta gttagctaac tagtacagac 2401 gcaggcgcaa acattagatg ccggcacaaa cacgcacaaa gacagagaca gacacgaaac 2461 gaccgctggc cagcttacct cccggcgggg gggtcggtgg tccctgggca ggggtctcca 2521 aatctcggtg gaacctccaa atgaaagacc cccgaggtgg gcagtcaatc aatctgagga 2581 gaccctccca aggatcagcg agtccacgat tcggatgcaa acagcaagag gctttattgg 2641 gaatacgggt acccgggcga cgcagtctat cggaggactg gcgcgccgag tgaggggttg 2701 tgggctcttt tattgagctc ggggagcaga agcgcgcgaa cagaagcgag aagcgaactg 2761 attggttagt tcaaataagg cacagggtca tttcaggtcc ttggggcacc ctggaaacat 2821 ctgatggttc tctagaaact gctgagggcg ggaccgcatc tggggaccat ctgttcttgg 2881 ccctgagccg gggcaggaac tgcttaccac agatatcctg tttggcccat attctgctgt 2941 ctctctgttc ctaaccttga tctgaacttc tctattctca gttatgtatt ttccatgcct 3001 tgcaaaatgg cgttacttaa gctagcttgc caaacctaca ggtggggtct ttcattcccc 3061 cctttttctg gagactaaat aaaatctttt attttatcta tggctcgtac tctataggct 3121 tcagctggtg atattgttga gtcaaaacta gagcctggac cactgatatc ctgtctttaa 3181 caaattggac taatcgcctc tacgccggac gcatcgtggc cggcatcacc ggcgccacag 3241 gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg gctcgccact 3301 tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg gccgggggac 3361 tgttgggcgc catctccttg catgcaccat tccttgcgcc gtcgttttac aacgtcgtga 3421 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag 3481 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa 3541 tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg 3601 catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca 3661 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag 3721 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa 3781 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat 3841 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg 3901 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 3961 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 4021 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 4081 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 4141 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 4201 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg 4261 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 4321 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 4381 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 4441 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 4501 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 4561 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 4621 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 4681 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 4741 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 4801 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 4861 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 4921 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 4981 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 5041 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 5101 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 5161 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 5221 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 5281 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 5341 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 5401 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 5461 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 5521 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 5581 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 5641 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 5701 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 5761 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc 5821 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag 5881 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt 5941 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acaattctcc 6001 ccagctcttt ctgcgctgtc cataatgaac tatttcagga gtattggagt atgtggattc 6061 aaaacagtgg ctaattgagt caggattact taatattcga ctcctaccct cagaatgttg 6121 ttctccaact tcaagaaact gttacccatg aaagaccccc gctgacgggt agtcaatcac 6181 tcagaggaga ccctcccaag gaacagcgag accacaagtc ggatgcaact gcaagagggt 6241 ttattggata cacgggtacc cgggcgacgc agtctatcgg aggactggcg cgccgagtga 6301 ggggttgtgg gctcttttat tgagctcggg gagcagaagc gcgcgaacag aagcgagaag 6361 cgaactgatt ggttagttca aataaggcac agggtcattt caggtccttg gggcaccctg 6421 gaaacatctg atggttctct agaaactgct gagggcggga ccgcatctgg ggaccatctg 6481 ttcttggccc tgagccgggg caggaactgc ttaccacaga tatcctgttt ggcccatatt 6541 ctgctgtctc tctgttccta accttgatct gaacttctct attctcagtt atgtattttc 6601 catgccttgc aaaatggcgt tacttaagct agcttgccaa acctacaggt ggggtctttc 6661 attcccccct ttttctggag actaaataaa atcttttatt ttatcgataa gcttggctgc 6721 aggtcgacgg tatcgataag cttgataatt cttaattaat tcattacttg tacagctcgt 6781 ccatgccgag agtgatcccg gcggcggtca cgaactccag caggaccatg tgatcgcgct 6841 tctcgttggg gtctttgctc agggcggact gggtgctcag gtagtggttg tcgggcagca 6901 gcacggggcc gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc tgcacgctgc 6961 cgtcctcgat gttgtggcgg atcttgaagt tcgccttgat gccgttcttc t //