LOCUS       sequence-307047-        3214 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Shuttle vector with mTurquoise2-tagged ITGB1 cytoplasmic tail as an 
            insert.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3214)
  AUTHORS   Miihkinen M, Gronloh MLB, Popovic A, Vihinen H, Jokitalo E, Goult 
            BT, Ivaska J, Jacquemet G
  TITLE     Myosin-X and talin modulate integrin activity at filopodia tips.
  JOURNAL   Cell Rep. 2021 Sep 14;36(11):109716. doi: 
            10.1016/j.celrep.2021.109716.
  PUBMED    34525374
REFERENCE   2  (bases 1 to 3214)
  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"; journalName: "Cell Rep.";
            date: "2021-09-14"; volume: "36(11):109716. doi"; pages: " 
            10.1016/j.celrep.2021.109716"
COMMENT     Sequence Label: pENTR2b ITGB1-tail-mTurquoise2
FEATURES             Location/Qualifiers
     source          1..3214
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      103..189
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      281..308
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     316..339
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    358..457
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      496..505
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(709..711,712..714,715..1428)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(754..775)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1015..1034)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1362..1383
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1443..1542)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1654..1673)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1665..2474
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1732..1751)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2564..3152
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3053..3072
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
       61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
      241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
      301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
      361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
      421 ttttttataa tgccaacttt gtacaaaaaa gcaggctggc gccggaacca attcagtcga
      481 ctggatccgg tacccgccac catggctcaa gcttcgaatt caggctgcat gagctgcaag
      541 tgtgtgctct ccaagctttt aatgataatt catgacagaa gggagtttgc taaatttgaa
      601 aaggagaaaa tgaatgccaa atgggacacg ggtgaaaatc ctatttataa gagtgccgta
      661 acaactgtgg tcaatccgaa gtatgaggga aaaggcggcg gcggccgcat ggtgagcaag
      721 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
      781 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
      841 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
      901 ctgtcctggg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
      961 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     1021 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     1081 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     1141 aactacttta gcgacaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc
     1201 aacttcaaga tccgccacaa catcgaggac ggcggcgtgc agctcgccga ccactaccag
     1261 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     1321 cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     1381 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaact cgagatatct
     1441 agacccagct ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg
     1501 caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag ctgcagctct
     1561 ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa
     1621 caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac
     1681 gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
     1741 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg
     1801 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
     1861 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
     1921 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc
     1981 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
     2041 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
     2101 gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
     2161 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat
     2221 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
     2281 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
     2341 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     2401 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
     2461 atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ttattcagat tgggccccgt
     2521 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     2581 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     2641 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     2701 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     2761 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     2821 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     2881 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     2941 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3001 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3061 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3121 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3181 tcctggcctt ttgctggcct tttgctcaca tgtt
//
