LOCUS       sequence-204239-        6719 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To study altered downstream functions of phospholylation-site null 
            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 S159/666A
FEATURES             Location/Qualifiers
     source          1..6719
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        54..357
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        358..561
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     512..532
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      591..600
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2658..2660,2661..2663,2664..3377)
                     /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(2658..2660,2661..2663,2664..3374)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2703..2724)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2964..2983)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3311..3332
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3500..3621
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3537..3556)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3591..3610
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3628..4083)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3765..3786)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3977..3996
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4110..4214
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4216..4573
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4216..4407
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4239..4259)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4424..4559
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4608..5402
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4608..5402
                     /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(4662..4681)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5272..5291
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5590..5609)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5634..5681
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6010..6598
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6499..6518
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
       61 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      121 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      181 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      241 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      301 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      361 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      421 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      481 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      541 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg gccaccatgg
      601 ctctggacgg gataaggatg ccagatggct gctacgcgga cgggacgtgg gaactgagtg
      661 tccatgtgac ggacctgaac cgcgatgtca ccctgagagt gaccggcgag gtgcacattg
      721 gaggcgtgat gcttaagctg gtggagaaac tcgatgtaaa aaaagattgg tctgaccatg
      781 ctctctggtg ggaaaagaag agaacttggc ttctgaagac acattggacc ttagataagt
      841 atggtattca ggcagatgct aagcttcagt tcacccctca gcacaaactg ctccgcctgc
      901 agcttcccaa catgaagtat gtgaaggtga aagtgaattt ctctgataga gtcttcaaag
      961 ctgtttctga catctgtaag acttttaata tcagacaccc cgaagaactt tctctcttaa
     1021 agaaacccag agatccaaca aagaaaaaaa agaagaagct agatgaccag gctgaagatg
     1081 aggcacttga attagagggg cctcttatca ctcctggatc aggaagtata tattcaagcc
     1141 caggactgta tagtaaaaca atgaccccca cttatgatgc tcatgatgga agccccttgt
     1201 caccaacttc tgcttggttt ggtgacagtg ctttgtcaga aggcaatcct ggtatacttg
     1261 ctgtcagtca accaatcacg tcaccagaaa tcttggcaaa aatgttcaag cctcaagctc
     1321 ttcttgataa agcaaaaatc aaccaaggat ggcttgattc ctcaagatct ctcatggaac
     1381 aagatgtgaa ggaaaatgag gccttgctgc tccgattcaa gtattacagc ttttttgatt
     1441 tgaatccaaa gtatgatgca atcagaatca atcagcttta tgagcaggcc aaatgggcca
     1501 ttctcctgga agagattgaa tgcacagaag aagaaatgat gatgtttgca gccctgcagt
     1561 atcatatcaa taagctgtca atcatgacat cagagaatca tttgaacaac agtgacaaag
     1621 aagttgatga agttgatgct gccctttcag acctggagat tactctggaa gggggtaaaa
     1681 cgtcaacaat tttgggtgac attacttcca ttcctgaact tgctgactac attaaagttt
     1741 tcaagccaaa aaagctgact ctgaaaggtt acaaacaata ttggtgcacc ttcaaagaca
     1801 catccatttc ttgttataag agcaaagaag aatccagtgg cacaccagct catcagatga
     1861 acctcagggg atgtgaagtt accccagatg taaacatttc aggccaaaaa tttaacatta
     1921 aactcctgat tccagttgca gaaggcatga atgaaatctg gcttcgttgt gacaatgaaa
     1981 aacagtatgc acactggatg gcagcctgca gattagcctc caaaggcaag accatggcgg
     2041 acagttctta caacttagaa gttcagaata ttctttcctt tctgaagatg cagcatttaa
     2101 acccagatcc tcagttaata ccagagcaga tcacgactga tataactcct gaatgtttgg
     2161 tgtctccccg ctatctaaaa aagtataaga acaagcagat aacagcgaga atcttggagg
     2221 cccatcagaa tgtagctcag atgagtctaa ttgaagccaa gatgagattt attcaagctt
     2281 ggcagtcact acctgaattt ggcatcactc acttcattgc aaggttccaa gggggcaaaa
     2341 aagaagaact tattggaatt gcatacaaca gactgattcg gatggatgcc agcactggag
     2401 atgcaattaa aacatggcgt ttcagcaaca tgaaacagtg gaatgtcaac tgggaaatca
     2461 aaatggtcac cgtagagttt gcagatgaag tacgattgtc cttcatttgt actgaagtag
     2521 attgcaaagt ggttcatgaa ttcattggtg gctacatatt tctctcaaca cgtgcaaaag
     2581 accaaaacga ggcattagat gaagagatgt tctacaaact taccagtggt tgggtgggta
     2641 actcagggga tccaggaatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     2701 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     2761 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     2821 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     2881 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     2941 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     3001 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     3061 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     3121 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     3181 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     3241 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     3301 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     3361 acgagctgta caagtaaagc ggccgcgact ctagatcata atcagccata ccacatttgt
     3421 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     3481 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     3541 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     3601 caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg
     3661 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     3721 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     3781 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     3841 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     3901 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     3961 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     4021 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     4081 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4141 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4201 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     4261 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4321 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4381 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     4441 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     4501 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     4561 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     4621 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     4681 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4741 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     4801 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4861 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     4921 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4981 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     5041 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     5101 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     5161 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     5221 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     5281 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     5341 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     5401 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     5461 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     5521 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
     5581 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     5641 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     5701 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     5761 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     5821 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
     5881 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     5941 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     6001 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     6061 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     6121 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     6181 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6241 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     6301 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     6361 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     6421 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     6481 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     6541 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     6601 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     6661 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttat
//
