LOCUS       sequence-204247-        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 S159/181/666E
FEATURES             Location/Qualifiers
     source          1..6719
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      30..618
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     519..538
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        793..1096
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1097..1300
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1251..1271
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1330..1339
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3397..3399,3400..3402,3403..4116)
                     /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(3397..3399,3400..3402,3403..4113)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3442..3463)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3703..3722)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4050..4071
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4239..4360
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4276..4295)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4330..4349
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4367..4822)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4504..4525)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4716..4735
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4849..4953
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4955..5312
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4955..5146
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4978..4998)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5163..5298
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5347..6141
                     /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(5401..5420)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6011..6030
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6329..6348)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6373..6420
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
       61 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
      121 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
      181 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
      241 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
      301 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
      361 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
      421 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
      481 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
      541 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
      601 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
      661 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
      721 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      781 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      841 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      901 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      961 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     1021 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     1081 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     1141 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     1201 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     1261 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
     1321 ccgctagcgg ccaccatggc tctggacggg ataaggatgc cagatggctg ctacgcggac
     1381 gggacgtggg aactgagtgt ccatgtgacg gacctgaacc gcgatgtcac cctgagagtg
     1441 accggcgagg tgcacattgg aggcgtgatg cttaagctgg tggagaaact cgatgtaaaa
     1501 aaagattggt ctgaccatgc tctctggtgg gaaaagaaga gaacttggct tctgaagaca
     1561 cattggacct tagataagta tggtattcag gcagatgcta agcttcagtt cacccctcag
     1621 cacaaactgc tccgcctgca gcttcccaac atgaagtatg tgaaggtgaa agtgaatttc
     1681 tctgatagag tcttcaaagc tgtttctgac atctgtaaga cttttaatat cagacacccc
     1741 gaagaacttt ctctcttaaa gaaacccaga gatccaacaa agaaaaaaaa gaagaagcta
     1801 gatgaccagg aggaagatga ggcacttgaa ttagaggggc ctcttatcac tcctggatca
     1861 ggaagtatat attcagagcc aggactgtat agtaaaacaa tgacccccac ttatgatgct
     1921 catgatggaa gccccttgtc accaacttct gcttggtttg gtgacagtgc tttgtcagaa
     1981 ggcaatcctg gtatacttgc tgtcagtcaa ccaatcacgt caccagaaat cttggcaaaa
     2041 atgttcaagc ctcaagctct tcttgataaa gcaaaaatca accaaggatg gcttgattcc
     2101 tcaagatctc tcatggaaca agatgtgaag gaaaatgagg ccttgctgct ccgattcaag
     2161 tattacagct tttttgattt gaatccaaag tatgatgcaa tcagaatcaa tcagctttat
     2221 gagcaggcca aatgggccat tctcctggaa gagattgaat gcacagaaga agaaatgatg
     2281 atgtttgcag ccctgcagta tcatatcaat aagctgtcaa tcatgacatc agagaatcat
     2341 ttgaacaaca gtgacaaaga agttgatgaa gttgatgctg ccctttcaga cctggagatt
     2401 actctggaag ggggtaaaac gtcaacaatt ttgggtgaca ttacttccat tcctgaactt
     2461 gctgactaca ttaaagtttt caagccaaaa aagctgactc tgaaaggtta caaacaatat
     2521 tggtgcacct tcaaagacac atccatttct tgttataaga gcaaagaaga atccagtggc
     2581 acaccagctc atcagatgaa cctcagggga tgtgaagtta ccccagatgt aaacatttca
     2641 ggccaaaaat ttaacattaa actcctgatt ccagttgcag aaggcatgaa tgaaatctgg
     2701 cttcgttgtg acaatgaaaa acagtatgca cactggatgg cagcctgcag attagcctcc
     2761 aaaggcaaga ccatggcgga cagttcttac aacttagaag ttcagaatat tctttccttt
     2821 ctgaagatgc agcatttaaa cccagatcct cagttaatac cagagcagat cacgactgat
     2881 ataactcctg aatgtttggt gtctccccgc tatctaaaaa agtataagaa caagcagata
     2941 acagcgagaa tcttggaggc ccatcagaat gtagctcaga tgagtctaat tgaagccaag
     3001 atgagattta ttcaagcttg gcagtcacta cctgaatttg gcatcactca cttcattgca
     3061 aggttccaag ggggcaaaaa agaagaactt attggaattg catacaacag actgattcgg
     3121 atggatgcca gcactggaga tgcaattaaa acatggcgtt tcagcaacat gaaacagtgg
     3181 aatgtcaact gggaaatcaa aatggtcacc gtagagtttg cagatgaagt acgattgtcc
     3241 ttcatttgta ctgaagtaga ttgcaaagtg gttcatgaat tcattggtgg ctacatattt
     3301 ctctcaacac gtgcaaaaga ccaaaacgag gaattagatg aagagatgtt ctacaaactt
     3361 accagtggtt gggtgggtaa ctcaggggat ccaggaatgg tgagcaaggg cgaggagctg
     3421 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     3481 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     3541 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     3601 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     3661 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     3721 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     3781 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     3841 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     3901 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     3961 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     4021 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     4081 gggatcactc tcggcatgga cgagctgtac aagtaaagcg gccgcgactc tagatcataa
     4141 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     4201 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     4261 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4321 attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt
     4381 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     4441 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     4501 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     4561 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     4621 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     4681 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     4741 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     4801 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4861 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4921 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     4981 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     5041 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     5101 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     5161 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     5221 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     5281 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     5341 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     5401 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     5461 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     5521 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     5581 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     5641 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     5701 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     5761 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     5821 gacaaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     5881 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     5941 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     6001 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     6061 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     6121 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     6181 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     6241 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     6301 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     6361 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     6421 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     6481 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     6541 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     6601 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     6661 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtca
//
