LOCUS Exported 7405 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS Flag-betaPIXa SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7405) AUTHORS Mayhew MW, Webb DJ, Kovalenko M, Whitmore L, Fox JW, Horwitz AF TITLE Identification of protein networks associated with the PAK1-betaPIX-GIT1-paxillin signaling complex by mass spectrometry. JOURNAL J Proteome Res. 2006 Sep . 5(9):2417-23. PUBMED 16944954 REFERENCE 2 (bases 1 to 7405) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7405 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 894..903 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 903..926 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" promoter complement(2958..2976) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(2959..2976) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(2996..3013) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3002..3226 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3272..3700 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3359..3378) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3569..3590 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3709..3729) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3714..4043 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3894..4029 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3956..3975 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4110..4904 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(4164..4183) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4774..4793 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5078..5199 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5115..5134) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5169..5188 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5248..5264) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5248..5264) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5261..5283) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5272..5288 /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(5296..5326) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5341..5362 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5479..5496) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5650..6238) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5730..5749) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6409..7269) /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 7032..7051 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7270..7374) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctgccacca 901 tggactacaa ggacgacgat gacaagggac ctaagcttgg taccgagctc ggatccacta 961 gtaacggccg ccagtgtgct ggaattcgca tgactgataa caccaacagc caactggtag 1021 tacgagccaa gtttaacttc cagcagacca atgaagatga actctccttc tcaaagggtg 1081 atgtcatcca tgtcacacga gtggaggaag gaggctggtg ggagggcaca cacaatggca 1141 ggaccggctg gttccccagc aactacgttc gagagatcaa gccaagtgag aagcccgtgt 1201 cacccaaatc agggaccttg aagagccctc ccaaagggtt cgatacgact gccatcaaca 1261 agagctatta caacgtggtg ctacagaaca tcctggaaac agagcatgag tattccaagg 1321 agctgcagtc tgtgctgtcc acctacctgc ggccactgca gaccagtgac aagttgagtt 1381 cagcaaacac ttcatattta atgggaaatc tagaggaaat atcttccttc cagcaagtgc 1441 ttgtacagtc cctagaagaa tgcacgaagt ctcctgaagc ccaacagaga gttggtggct 1501 gcttcctgag cctgatgccg cagatgagga ccctgtacct cgcttactgt gccaaccacc 1561 catctgctgt gagcgtcctc acagagcaca gtgaggacct aggagagttc atggaaacaa 1621 aaggtgccag cagccctggg atcctggtgc tgaccaccgg cctgagcaag cccttcatgc 1681 gcctggacaa gtaccccaca ctgctaaagg agctggagag acacatggag gattatcatc 1741 ctgatagaca agatattcag aagtctatga cggccttcaa aaacctttca gctcagtgtc 1801 aagaagttcg caagaggaag gagctggagc tgcagatcct gacggagccc atcaggagct 1861 gggaggggga tgacataaag accctgggca gtgtcacata catgtcccaa gtcaccattc 1921 agtgtgcggg aagcgaggag aagaatgaga gatacctcct gctcttccca aaccttctgc 1981 tcatgttgtc tgcaagtccc aggatgagtg gtttcatcta tcaggggaag ctgccaacaa 2041 caggaatgac aatcacaaag cttgaggaca gtgaaaacca taggaatgca tttgagatat 2101 cagggagcat gatcgagcgg attctggtgt cctgcaccag ccagcaggac ttacacgagt 2161 gggtggaaca cctgcagaag cagacgaagg tcacatctgt gagcaacccc accatcaaac 2221 cccactcggt gccatcacac acacttcctt cccatcctct cactccatcc agcaaacacg 2281 cggacagcaa gcccgtggca ctgacgcctg cgtaccacac actcccccac ccctctcacc 2341 atggcacccc acacaccacc atcagctggg gacccctgga gcctccgaag acccccaagc 2401 cttggagcct gagttgcctg cggcctgcac ctcccctccg gccctcagct gctctctgct 2461 acaaggagga tctcagtaag agccccaaga ccatgaaaaa gctgctgccg aagcgcaagc 2521 ccgagcggaa gccttcggac gaggagttcg ctgtgcgcaa gagcacagcg gcgctggaag 2581 aagacgctca gatcctgaag gttatcgaag cttattgcac aagtgcaaag acgcgccaga 2641 ccctgaactc aagttcacgc aaagaatctg ctccacaagt gttgcttcca gaagaagaaa 2701 aaattatagt cgaagaaacc aagagcaatg ggcagacagt gatagaggaa aagagcctcg 2761 tggatacggt gtatgcgtta aaggatgaag tccaggagtt aagacaggat aacaagaaga 2821 tgaagaagtc cttagaggag gaacagaggg cccgcaagga cttagagaag ctggtgagga 2881 aggttctaaa gaatatgaac gaccctgcct gggatgagac caatctatag ctcgagcatg 2941 catctagagg gccctattct atagtgtcac ctaaatgcta gagctcgctg atcagcctcg 3001 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc 3061 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt 3121 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat 3181 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa 3241 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg 3301 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct 3361 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta 3421 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa 3481 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct 3541 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc 3601 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg 3661 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc 3721 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc 3781 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 3841 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc 3901 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt 3961 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt 4021 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg 4081 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt 4141 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct 4201 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga 4261 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg 4321 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact 4381 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg 4441 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct 4501 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg 4561 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt 4621 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg 4681 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc 4741 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag 4801 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt 4861 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt 4921 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc 4981 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca 5041 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa 5101 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca 5161 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac 5221 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc 5281 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 5341 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 5401 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 5461 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 5521 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 5581 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 5641 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 5701 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 5761 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 5821 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 5881 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 5941 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 6001 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 6061 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa 6121 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 6181 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 6241 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 6301 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 6361 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 6421 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 6481 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 6541 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 6601 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 6661 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 6721 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 6781 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 6841 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 6901 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 6961 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 7021 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 7081 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 7141 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 7201 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 7261 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 7321 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 7381 tccccgaaaa gtgccacctg acgtc //