LOCUS       sequence-204242-        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 S159E
FEATURES             Location/Qualifiers
     source          1..6719
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      15..603
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     504..523
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        778..1081
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1082..1285
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1236..1256
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1315..1324
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3382..3384,3385..3387,3388..4101)
                     /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(3382..3384,3385..3387,3388..4098)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3427..3448)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3688..3707)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4035..4056
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4224..4345
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4261..4280)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4315..4334
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4352..4807)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4489..4510)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4701..4720
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4834..4938
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4940..5297
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4940..5131
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4963..4983)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5148..5283
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5332..6126
                     /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(5386..5405)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5996..6015
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6314..6333)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6358..6405
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
       61 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
      121 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
      181 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
      241 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
      301 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
      361 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
      421 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
      481 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
      541 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
      601 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
      661 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag
      721 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      781 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      841 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      901 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      961 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     1021 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     1081 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     1141 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     1201 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     1261 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agcggccacc
     1321 atggctctgg acgggataag gatgccagat ggctgctacg cggacgggac gtgggaactg
     1381 agtgtccatg tgacggacct gaaccgcgat gtcaccctga gagtgaccgg cgaggtgcac
     1441 attggaggcg tgatgcttaa gctggtggag aaactcgatg taaaaaaaga ttggtctgac
     1501 catgctctct ggtgggaaaa gaagagaact tggcttctga agacacattg gaccttagat
     1561 aagtatggta ttcaggcaga tgctaagctt cagttcaccc ctcagcacaa actgctccgc
     1621 ctgcagcttc ccaacatgaa gtatgtgaag gtgaaagtga atttctctga tagagtcttc
     1681 aaagctgttt ctgacatctg taagactttt aatatcagac accccgaaga actttctctc
     1741 ttaaagaaac ccagagatcc aacaaagaaa aaaaagaaga agctagatga ccaggaggaa
     1801 gatgaggcac ttgaattaga ggggcctctt atcactcctg gatcaggaag tatatattca
     1861 agcccaggac tgtatagtaa aacaatgacc cccacttatg atgctcatga tggaagcccc
     1921 ttgtcaccaa cttctgcttg gtttggtgac agtgctttgt cagaaggcaa tcctggtata
     1981 cttgctgtca gtcaaccaat cacgtcacca gaaatcttgg caaaaatgtt caagcctcaa
     2041 gctcttcttg ataaagcaaa aatcaaccaa ggatggcttg attcctcaag atctctcatg
     2101 gaacaagatg tgaaggaaaa tgaggccttg ctgctccgat tcaagtatta cagctttttt
     2161 gatttgaatc caaagtatga tgcaatcaga atcaatcagc tttatgagca ggccaaatgg
     2221 gccattctcc tggaagagat tgaatgcaca gaagaagaaa tgatgatgtt tgcagccctg
     2281 cagtatcata tcaataagct gtcaatcatg acatcagaga atcatttgaa caacagtgac
     2341 aaagaagttg atgaagttga tgctgccctt tcagacctgg agattactct ggaagggggt
     2401 aaaacgtcaa caattttggg tgacattact tccattcctg aacttgctga ctacattaaa
     2461 gttttcaagc caaaaaagct gactctgaaa ggttacaaac aatattggtg caccttcaaa
     2521 gacacatcca tttcttgtta taagagcaaa gaagaatcca gtggcacacc agctcatcag
     2581 atgaacctca ggggatgtga agttacccca gatgtaaaca tttcaggcca aaaatttaac
     2641 attaaactcc tgattccagt tgcagaaggc atgaatgaaa tctggcttcg ttgtgacaat
     2701 gaaaaacagt atgcacactg gatggcagcc tgcagattag cctccaaagg caagaccatg
     2761 gcggacagtt cttacaactt agaagttcag aatattcttt cctttctgaa gatgcagcat
     2821 ttaaacccag atcctcagtt aataccagag cagatcacga ctgatataac tcctgaatgt
     2881 ttggtgtctc cccgctatct aaaaaagtat aagaacaagc agataacagc gagaatcttg
     2941 gaggcccatc agaatgtagc tcagatgagt ctaattgaag ccaagatgag atttattcaa
     3001 gcttggcagt cactacctga atttggcatc actcacttca ttgcaaggtt ccaagggggc
     3061 aaaaaagaag aacttattgg aattgcatac aacagactga ttcggatgga tgccagcact
     3121 ggagatgcaa ttaaaacatg gcgtttcagc aacatgaaac agtggaatgt caactgggaa
     3181 atcaaaatgg tcaccgtaga gtttgcagat gaagtacgat tgtccttcat ttgtactgaa
     3241 gtagattgca aagtggttca tgaattcatt ggtggctaca tatttctctc aacacgtgca
     3301 aaagaccaaa acgagagttt agatgaagag atgttctaca aacttaccag tggttgggtg
     3361 ggtaactcag gggatccagg aatggtgagc aagggcgagg agctgttcac cggggtggtg
     3421 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     3481 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     3541 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     3601 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     3661 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     3721 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     3781 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     3841 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     3901 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     3961 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     4021 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     4081 atggacgagc tgtacaagta aagcggccgc gactctagat cataatcagc cataccacat
     4141 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     4201 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     4261 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     4321 tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa
     4381 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     4441 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     4501 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     4561 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     4621 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     4681 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     4741 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     4801 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4861 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4921 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     4981 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     5041 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     5101 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     5161 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     5221 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     5281 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     5341 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     5401 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     5461 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     5521 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     5581 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     5641 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     5701 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     5761 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacaa agagcatcag
     5821 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     5881 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5941 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     6001 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     6061 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     6121 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     6181 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     6241 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     6301 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     6361 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     6421 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     6481 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     6541 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     6601 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     6661 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaag
//
