LOCUS       sequence-204245-        6719 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To study altered downstream functions of phospholylation-site 
            mimetic kindlin2.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1083/jcb"; 
            journalName: "J Cell Biol"; date: "2015-02-2- 2"; volume: "208"; 
            issue: "3"; pages: "331-50"
COMMENT     Sequence Label: pEGFP Kind2 S159/666E
FEATURES             Location/Qualifiers
     source          1..6719
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      18..606
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     507..526
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        781..1084
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1085..1288
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1239..1259
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1318..1327
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3385..3387,3388..3390,3391..4104)
                     /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             join(3385..3387,3388..3390,3391..4101)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3430..3451)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3691..3710)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4038..4059
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4227..4348
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4264..4283)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4318..4337
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4355..4810)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4492..4513)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4704..4723
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4837..4941
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4943..5300
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4943..5134
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4966..4986)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5151..5286
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5335..6129
                     /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
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDKEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5389..5408)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5999..6018
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6317..6336)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6361..6408
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
       61 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
      121 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
      181 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
      241 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
      301 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
      361 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
      421 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
      481 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
      541 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
      601 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
      661 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
      721 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
      781 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      841 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      901 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      961 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     1021 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     1081 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     1141 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     1201 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     1261 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcggcc
     1321 accatggctc tggacgggat aaggatgcca gatggctgct acgcggacgg gacgtgggaa
     1381 ctgagtgtcc atgtgacgga cctgaaccgc gatgtcaccc tgagagtgac cggcgaggtg
     1441 cacattggag gcgtgatgct taagctggtg gagaaactcg atgtaaaaaa agattggtct
     1501 gaccatgctc tctggtggga aaagaagaga acttggcttc tgaagacaca ttggacctta
     1561 gataagtatg gtattcaggc agatgctaag cttcagttca cccctcagca caaactgctc
     1621 cgcctgcagc ttcccaacat gaagtatgtg aaggtgaaag tgaatttctc tgatagagtc
     1681 ttcaaagctg tttctgacat ctgtaagact tttaatatca gacaccccga agaactttct
     1741 ctcttaaaga aacccagaga tccaacaaag aaaaaaaaga agaagctaga tgaccaggag
     1801 gaagatgagg cacttgaatt agaggggcct cttatcactc ctggatcagg aagtatatat
     1861 tcaagcccag gactgtatag taaaacaatg acccccactt atgatgctca tgatggaagc
     1921 cccttgtcac caacttctgc ttggtttggt gacagtgctt tgtcagaagg caatcctggt
     1981 atacttgctg tcagtcaacc aatcacgtca ccagaaatct tggcaaaaat gttcaagcct
     2041 caagctcttc ttgataaagc aaaaatcaac caaggatggc ttgattcctc aagatctctc
     2101 atggaacaag atgtgaagga aaatgaggcc ttgctgctcc gattcaagta ttacagcttt
     2161 tttgatttga atccaaagta tgatgcaatc agaatcaatc agctttatga gcaggccaaa
     2221 tgggccattc tcctggaaga gattgaatgc acagaagaag aaatgatgat gtttgcagcc
     2281 ctgcagtatc atatcaataa gctgtcaatc atgacatcag agaatcattt gaacaacagt
     2341 gacaaagaag ttgatgaagt tgatgctgcc ctttcagacc tggagattac tctggaaggg
     2401 ggtaaaacgt caacaatttt gggtgacatt acttccattc ctgaacttgc tgactacatt
     2461 aaagttttca agccaaaaaa gctgactctg aaaggttaca aacaatattg gtgcaccttc
     2521 aaagacacat ccatttcttg ttataagagc aaagaagaat ccagtggcac accagctcat
     2581 cagatgaacc tcaggggatg tgaagttacc ccagatgtaa acatttcagg ccaaaaattt
     2641 aacattaaac tcctgattcc agttgcagaa ggcatgaatg aaatctggct tcgttgtgac
     2701 aatgaaaaac agtatgcaca ctggatggca gcctgcagat tagcctccaa aggcaagacc
     2761 atggcggaca gttcttacaa cttagaagtt cagaatattc tttcctttct gaagatgcag
     2821 catttaaacc cagatcctca gttaatacca gagcagatca cgactgatat aactcctgaa
     2881 tgtttggtgt ctccccgcta tctaaaaaag tataagaaca agcagataac agcgagaatc
     2941 ttggaggccc atcagaatgt agctcagatg agtctaattg aagccaagat gagatttatt
     3001 caagcttggc agtcactacc tgaatttggc atcactcact tcattgcaag gttccaaggg
     3061 ggcaaaaaag aagaacttat tggaattgca tacaacagac tgattcggat ggatgccagc
     3121 actggagatg caattaaaac atggcgtttc agcaacatga aacagtggaa tgtcaactgg
     3181 gaaatcaaaa tggtcaccgt agagtttgca gatgaagtac gattgtcctt catttgtact
     3241 gaagtagatt gcaaagtggt tcatgaattc attggtggct acatatttct ctcaacacgt
     3301 gcaaaagacc aaaacgagga attagatgaa gagatgttct acaaacttac cagtggttgg
     3361 gtgggtaact caggggatcc aggaatggtg agcaagggcg aggagctgtt caccggggtg
     3421 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     3481 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     3541 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     3601 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     3661 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     3721 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     3781 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     3841 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     3901 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     3961 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     4021 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     4081 ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gatcataatc agccatacca
     4141 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     4201 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     4261 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4321 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     4381 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     4441 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     4501 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     4561 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     4621 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     4681 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     4741 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     4801 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4861 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4921 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     4981 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     5041 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     5101 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     5161 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     5221 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     5281 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     5341 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     5401 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     5461 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     5521 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     5581 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     5641 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     5701 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     5761 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga caaagagcat
     5821 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     5881 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     5941 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     6001 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     6061 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     6121 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     6181 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     6241 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     6301 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     6361 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     6421 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     6481 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     6541 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     6601 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6661 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaa
//
