LOCUS       Exported                6719 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  To study effect of Kindlin 2 on Cell/ECM adhesion.
ACCESSION   .
VERSION     .
KEYWORDS    pEGFP Kindlin2
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 12, 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      10..598
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     499..518
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        773..1076
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1077..1280
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1231..1251
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1310..1319
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3377..4096
                     /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             3377..4093
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3422..3443)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3683..3702)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4030..4051
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4219..4340
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4256..4275)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4310..4329
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4347..4802)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4484..4505)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4696..4715
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4829..4933
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4935..5292
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(4958..4978)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5143..5278
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5205..5224
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5327..6121
                     /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(5381..5400)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5991..6010
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6309..6328)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6353..6400
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
       61 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
      121 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
      181 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
      241 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
      301 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
      361 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
      421 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
      481 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
      541 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
      601 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
      661 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt
      721 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
      781 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      841 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      901 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      961 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     1021 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
     1081 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     1141 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     1201 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     1261 aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgg ccaccatggc
     1321 tctggacggg ataaggatgc cagatggctg ctacgcggac gggacgtggg aactgagtgt
     1381 ccatgtgacg gacctgaacc gcgatgtcac cctgagagtg accggcgagg tgcacattgg
     1441 aggcgtgatg cttaagctgg tggagaaact cgatgtaaaa aaagattggt ctgaccatgc
     1501 tctctggtgg gaaaagaaga gaacttggct tctgaagaca cattggacct tagataagta
     1561 tggtattcag gcagatgcta agcttcagtt cacccctcag cacaaactgc tccgcctgca
     1621 gcttcccaac atgaagtatg tgaaggtgaa agtgaatttc tctgatagag tcttcaaagc
     1681 tgtttctgac atctgtaaga cttttaatat cagacacccc gaagaacttt ctctcttaaa
     1741 gaaacccaga gatccaacaa agaaaaaaaa gaagaagcta gatgaccagt ctgaagatga
     1801 ggcacttgaa ttagaggggc ctcttatcac tcctggatca ggaagtatat attcaagccc
     1861 aggactgtat agtaaaacaa tgacccccac ttatgatgct catgatggaa gccccttgtc
     1921 accaacttct gcttggtttg gtgacagtgc tttgtcagaa ggcaatcctg gtatacttgc
     1981 tgtcagtcaa ccaatcacgt caccagaaat cttggcaaaa atgttcaagc ctcaagctct
     2041 tcttgataaa gcaaaaatca accaaggatg gcttgattcc tcaagatctc tcatggaaca
     2101 agatgtgaag gaaaatgagg ccttgctgct ccgattcaag tattacagct tttttgattt
     2161 gaatccaaag tatgatgcaa tcagaatcaa tcagctttat gagcaggcca aatgggccat
     2221 tctcctggaa gagattgaat gcacagaaga agaaatgatg atgtttgcag ccctgcagta
     2281 tcatatcaat aagctgtcaa tcatgacatc agagaatcat ttgaacaaca gtgacaaaga
     2341 agttgatgaa gttgatgctg ccctttcaga cctggagatt actctggaag ggggtaaaac
     2401 gtcaacaatt ttgggtgaca ttacttccat tcctgaactt gctgactaca ttaaagtttt
     2461 caagccaaaa aagctgactc tgaaaggtta caaacaatat tggtgcacct tcaaagacac
     2521 atccatttct tgttataaga gcaaagaaga atccagtggc acaccagctc atcagatgaa
     2581 cctcagggga tgtgaagtta ccccagatgt aaacatttca ggccaaaaat ttaacattaa
     2641 actcctgatt ccagttgcag aaggcatgaa tgaaatctgg cttcgttgtg acaatgaaaa
     2701 acagtatgca cactggatgg cagcctgcag attagcctcc aaaggcaaga ccatggcgga
     2761 cagttcttac aacttagaag ttcagaatat tctttccttt ctgaagatgc agcatttaaa
     2821 cccagatcct cagttaatac cagagcagat cacgactgat ataactcctg aatgtttggt
     2881 gtctccccgc tatctaaaaa agtataagaa caagcagata acagcgagaa tcttggaggc
     2941 ccatcagaat gtagctcaga tgagtctaat tgaagccaag atgagattta ttcaagcttg
     3001 gcagtcacta cctgaatttg gcatcactca cttcattgca aggttccaag ggggcaaaaa
     3061 agaagaactt attggaattg catacaacag actgattcgg atggatgcca gcactggaga
     3121 tgcaattaaa acatggcgtt tcagcaacat gaaacagtgg aatgtcaact gggaaatcaa
     3181 aatggtcacc gtagagtttg cagatgaagt acgattgtcc ttcatttgta ctgaagtaga
     3241 ttgcaaagtg gttcatgaat tcattggtgg ctacatattt ctctcaacac gtgcaaaaga
     3301 ccaaaacgag agtttagatg aagagatgtt ctacaaactt accagtggtt gggtgggtaa
     3361 ctcaggggat ccaggaatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     3421 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     3481 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     3541 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     3601 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     3661 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     3721 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     3781 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     3841 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     3901 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     3961 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     4021 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     4081 cgagctgtac aagtaaagcg gccgcgactc tagatcataa tcagccatac cacatttgta
     4141 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     4201 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     4261 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     4321 aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     4381 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     4441 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     4501 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     4561 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     4621 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     4681 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     4741 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     4801 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     4861 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4921 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     4981 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5041 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5101 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     5161 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     5221 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     5281 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     5341 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     5401 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     5461 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     5521 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     5581 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     5641 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     5701 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     5761 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     5821 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     5881 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     5941 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     6001 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     6061 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     6121 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     6181 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     6241 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     6301 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     6361 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     6421 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     6481 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     6541 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     6601 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6661 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaa
//
