LOCUS Exported 6719 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION To study altered downstream functions of phospholylation-site null kindlin2. ACCESSION . VERSION . KEYWORDS pEGFP KInd2 S159A SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6719) AUTHORS Artym VV, Swatkoski S, Matsumoto K, Campbell CB, Petrie RJ, Dimitriadis EK, Li X, Mueller SC, Bugge TH, Gucek M, Yamada KM TITLE Dense fibrillar collagen is a potent inducer of invadopodia via a specific signaling network. JOURNAL J Cell Biol. 2015 Feb 2;208(3):331-50. doi: 10.1083/jcb.201405099. PUBMED 25646088 REFERENCE 2 (bases 1 to 6719) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6719 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 20..608 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 509..528 /label=pBR322ori-F /note="pBR322 origin, forward primer" enhancer 783..1086 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 1087..1290 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 1241..1261 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 1320..1329 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3387..4106 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 3387..4103 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3432..3453) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3693..3712) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4040..4061 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 4229..4350 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4266..4285) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4320..4339 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(4357..4812) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4494..4515) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 4706..4725 /label=F1ori-R /note="F1 origin, reverse primer" promoter 4839..4943 /gene="bla" /label=AmpR promoter promoter 4945..5302 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(4968..4988) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 5153..5288 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5215..5234 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5337..6131 /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 GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(5391..5410) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6001..6020 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(6319..6338) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 6363..6410 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" ORIGIN 1 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa 61 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt 121 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc 181 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa 241 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa 301 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc 361 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa 421 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa 481 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg 541 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc 601 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg 661 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc 721 attagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc 781 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca 841 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt 901 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg 961 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag 1021 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt 1081 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg 1141 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa 1201 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt 1261 gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgg 1321 ccaccatggc tctggacggg ataaggatgc cagatggctg ctacgcggac gggacgtggg 1381 aactgagtgt ccatgtgacg gacctgaacc gcgatgtcac cctgagagtg accggcgagg 1441 tgcacattgg aggcgtgatg cttaagctgg tggagaaact cgatgtaaaa aaagattggt 1501 ctgaccatgc tctctggtgg gaaaagaaga gaacttggct tctgaagaca cattggacct 1561 tagataagta tggtattcag gcagatgcta agcttcagtt cacccctcag cacaaactgc 1621 tccgcctgca gcttcccaac atgaagtatg tgaaggtgaa agtgaatttc tctgatagag 1681 tcttcaaagc tgtttctgac atctgtaaga cttttaatat cagacacccc gaagaacttt 1741 ctctcttaaa gaaacccaga gatccaacaa agaaaaaaaa gaagaagcta gatgaccagg 1801 ctgaagatga ggcacttgaa ttagaggggc ctcttatcac tcctggatca ggaagtatat 1861 attcaagccc aggactgtat agtaaaacaa tgacccccac ttatgatgct catgatggaa 1921 gccccttgtc accaacttct gcttggtttg gtgacagtgc tttgtcagaa ggcaatcctg 1981 gtatacttgc tgtcagtcaa ccaatcacgt caccagaaat cttggcaaaa atgttcaagc 2041 ctcaagctct tcttgataaa gcaaaaatca accaaggatg gcttgattcc tcaagatctc 2101 tcatggaaca agatgtgaag gaaaatgagg ccttgctgct ccgattcaag tattacagct 2161 tttttgattt gaatccaaag tatgatgcaa tcagaatcaa tcagctttat gagcaggcca 2221 aatgggccat tctcctggaa gagattgaat gcacagaaga agaaatgatg atgtttgcag 2281 ccctgcagta tcatatcaat aagctgtcaa tcatgacatc agagaatcat ttgaacaaca 2341 gtgacaaaga agttgatgaa gttgatgctg ccctttcaga cctggagatt actctggaag 2401 ggggtaaaac gtcaacaatt ttgggtgaca ttacttccat tcctgaactt gctgactaca 2461 ttaaagtttt caagccaaaa aagctgactc tgaaaggtta caaacaatat tggtgcacct 2521 tcaaagacac atccatttct tgttataaga gcaaagaaga atccagtggc acaccagctc 2581 atcagatgaa cctcagggga tgtgaagtta ccccagatgt aaacatttca ggccaaaaat 2641 ttaacattaa actcctgatt ccagttgcag aaggcatgaa tgaaatctgg cttcgttgtg 2701 acaatgaaaa acagtatgca cactggatgg cagcctgcag attagcctcc aaaggcaaga 2761 ccatggcgga cagttcttac aacttagaag ttcagaatat tctttccttt ctgaagatgc 2821 agcatttaaa cccagatcct cagttaatac cagagcagat cacgactgat ataactcctg 2881 aatgtttggt gtctccccgc tatctaaaaa agtataagaa caagcagata acagcgagaa 2941 tcttggaggc ccatcagaat gtagctcaga tgagtctaat tgaagccaag atgagattta 3001 ttcaagcttg gcagtcacta cctgaatttg gcatcactca cttcattgca aggttccaag 3061 ggggcaaaaa agaagaactt attggaattg catacaacag actgattcgg atggatgcca 3121 gcactggaga tgcaattaaa acatggcgtt tcagcaacat gaaacagtgg aatgtcaact 3181 gggaaatcaa aatggtcacc gtagagtttg cagatgaagt acgattgtcc ttcatttgta 3241 ctgaagtaga ttgcaaagtg gttcatgaat tcattggtgg ctacatattt ctctcaacac 3301 gtgcaaaaga ccaaaacgag agtttagatg aagagatgtt ctacaaactt accagtggtt 3361 gggtgggtaa ctcaggggat ccaggaatgg tgagcaaggg cgaggagctg ttcaccgggg 3421 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg 3481 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg 3541 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct 3601 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag 3661 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg 3721 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca 3781 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct 3841 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca 3901 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg 3961 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc 4021 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc 4081 tcggcatgga cgagctgtac aagtaaagcg gccgcgactc tagatcataa tcagccatac 4141 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa 4201 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa 4261 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg 4321 tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt 4381 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg 4441 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt 4501 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct 4561 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt 4621 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa 4681 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc 4741 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc 4801 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat 4861 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc 4921 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca 4981 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct 5041 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca 5101 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc 5161 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta 5221 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt 5281 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga 5341 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct 5401 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc 5461 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag 5521 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg 5581 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc 5641 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc 5701 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg 5761 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc 5821 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg 5881 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc 5941 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag 6001 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg 6061 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg 6121 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc 6181 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt 6241 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca 6301 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat 6361 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg 6421 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata 6481 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct 6541 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta 6601 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa 6661 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtag //