LOCUS       sequence-204236-        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 S666A
FEATURES             Location/Qualifiers
     source          1..6719
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      13..601
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     502..521
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        776..1079
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1080..1283
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1234..1254
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1313..1322
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3380..3382,3383..3385,3386..4099)
                     /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(3380..3382,3383..3385,3386..4096)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3425..3446)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3686..3705)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4033..4054
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4222..4343
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4259..4278)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4313..4332
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4350..4805)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4487..4508)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4699..4718
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4832..4936
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4938..5295
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4938..5129
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4961..4981)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5146..5281
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5330..6124
                     /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             5330..6124
                     /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(5384..5403)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5994..6013
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6312..6331)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6356..6403
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
       61 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
      121 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
      181 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
      241 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
      301 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
      361 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
      421 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
      481 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
      541 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
      601 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
      661 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt
      721 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
      781 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      841 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      901 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      961 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     1021 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     1081 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     1141 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     1201 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     1261 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag cggccaccat
     1321 ggctctggac gggataagga tgccagatgg ctgctacgcg gacgggacgt gggaactgag
     1381 tgtccatgtg acggacctga accgcgatgt caccctgaga gtgaccggcg aggtgcacat
     1441 tggaggcgtg atgcttaagc tggtggagaa actcgatgta aaaaaagatt ggtctgacca
     1501 tgctctctgg tgggaaaaga agagaacttg gcttctgaag acacattgga ccttagataa
     1561 gtatggtatt caggcagatg ctaagcttca gttcacccct cagcacaaac tgctccgcct
     1621 gcagcttccc aacatgaagt atgtgaaggt gaaagtgaat ttctctgata gagtcttcaa
     1681 agctgtttct gacatctgta agacttttaa tatcagacac cccgaagaac tttctctctt
     1741 aaagaaaccc agagatccaa caaagaaaaa aaagaagaag ctagatgacc agtctgaaga
     1801 tgaggcactt gaattagagg ggcctcttat cactcctgga tcaggaagta tatattcaag
     1861 cccaggactg tatagtaaaa caatgacccc cacttatgat gctcatgatg gaagcccctt
     1921 gtcaccaact tctgcttggt ttggtgacag tgctttgtca gaaggcaatc ctggtatact
     1981 tgctgtcagt caaccaatca cgtcaccaga aatcttggca aaaatgttca agcctcaagc
     2041 tcttcttgat aaagcaaaaa tcaaccaagg atggcttgat tcctcaagat ctctcatgga
     2101 acaagatgtg aaggaaaatg aggccttgct gctccgattc aagtattaca gcttttttga
     2161 tttgaatcca aagtatgatg caatcagaat caatcagctt tatgagcagg ccaaatgggc
     2221 cattctcctg gaagagattg aatgcacaga agaagaaatg atgatgtttg cagccctgca
     2281 gtatcatatc aataagctgt caatcatgac atcagagaat catttgaaca acagtgacaa
     2341 agaagttgat gaagttgatg ctgccctttc agacctggag attactctgg aagggggtaa
     2401 aacgtcaaca attttgggtg acattacttc cattcctgaa cttgctgact acattaaagt
     2461 tttcaagcca aaaaagctga ctctgaaagg ttacaaacaa tattggtgca ccttcaaaga
     2521 cacatccatt tcttgttata agagcaaaga agaatccagt ggcacaccag ctcatcagat
     2581 gaacctcagg ggatgtgaag ttaccccaga tgtaaacatt tcaggccaaa aatttaacat
     2641 taaactcctg attccagttg cagaaggcat gaatgaaatc tggcttcgtt gtgacaatga
     2701 aaaacagtat gcacactgga tggcagcctg cagattagcc tccaaaggca agaccatggc
     2761 ggacagttct tacaacttag aagttcagaa tattctttcc tttctgaaga tgcagcattt
     2821 aaacccagat cctcagttaa taccagagca gatcacgact gatataactc ctgaatgttt
     2881 ggtgtctccc cgctatctaa aaaagtataa gaacaagcag ataacagcga gaatcttgga
     2941 ggcccatcag aatgtagctc agatgagtct aattgaagcc aagatgagat ttattcaagc
     3001 ttggcagtca ctacctgaat ttggcatcac tcacttcatt gcaaggttcc aagggggcaa
     3061 aaaagaagaa cttattggaa ttgcatacaa cagactgatt cggatggatg ccagcactgg
     3121 agatgcaatt aaaacatggc gtttcagcaa catgaaacag tggaatgtca actgggaaat
     3181 caaaatggtc accgtagagt ttgcagatga agtacgattg tccttcattt gtactgaagt
     3241 agattgcaaa gtggttcatg aattcattgg tggctacata tttctctcaa cacgtgcaaa
     3301 agaccaaaac gaggcattag atgaagagat gttctacaaa cttaccagtg gttgggtggg
     3361 taactcaggg gatccaggaa tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     3421 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     3481 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     3541 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     3601 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     3661 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     3721 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     3781 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     3841 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     3901 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     3961 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     4021 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     4081 ggacgagctg tacaagtaaa gcggccgcga ctctagatca taatcagcca taccacattt
     4141 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     4201 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     4261 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4321 tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt
     4381 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     4441 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     4501 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     4561 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     4621 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     4681 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     4741 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     4801 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4861 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4921 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     4981 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     5041 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     5101 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     5161 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     5221 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     5281 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     5341 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     5401 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     5461 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     5521 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     5581 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     5641 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     5701 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     5761 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     5821 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     5881 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     5941 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     6001 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     6061 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     6121 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     6181 gatttcgatt ccaccgccgc cttctatgaa aggttggaca tcggaatcgt tttccgggac
     6241 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     6301 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     6361 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     6421 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     6481 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     6541 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     6601 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     6661 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagat
//
