LOCUS       sequence-471961-        5689 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector with VPS35 tagged with mScarlet.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5689)
  AUTHORS   Conway JRW, Joshi O, Kaivola J, Follain G, Gounis M, Kuhl D, Ivaska 
            J
  TITLE     Dynamic regulation of integrin beta1 phosphorylation supports 
            invasion of breast cancer cells.
  JOURNAL   Nat Cell Biol. 2025 May 26. doi: 10.1038/s41556-025-01663-4.
  PUBMED    40419795
REFERENCE   2  (bases 1 to 5689)
  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: "Nat Cell 
            Biol. 2025 May 26. doi: 10.1038/s41556-025-01663-4."
COMMENT     Sequence Label: pENTR221-VPS35-mScarlet
FEATURES             Location/Qualifiers
     source          1..5689
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3103..3798
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(3236..3254)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3504..3523
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(3814..3913)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3930..3949)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3931..3949)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3954..3970)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3954..3970)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4072..4091)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4083..4892
                     /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(4150..4169)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5039..5627
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5528..5547
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacacattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctgg cgccggaacc aattcagtcg actggtaccg gcaccatgcc tacaacacag
      721 cagtcccctc aggatgagca ggaaaagctc ttggatgaag ccatacaggc tgtgaaggtc
      781 cagtcattcc aaatgaagag atgcctggac aaaaacaagc ttatggatgc tctaaaacat
      841 gcttctaata tgcttggtga actccggact tctatgttat caccaaagag ttactatgaa
      901 ctttatatgg ccatttctga tgaactgcac tacttggagg tctacctgac agatgagttt
      961 gctaaaggaa ggaaagtggc agatctctac gaacttgtac agtatgctgg aaacattatc
     1021 ccaaggcttt accttttgat cacagttgga gttgtatatg tcaagtcatt tcctcagtcc
     1081 aggaaggata ttttgaaaga tttggtagaa atgtgccgtg gtgtgcaaca tcccttgagg
     1141 ggtctgtttc ttcgaaatta ccttcttcag tgtaccagaa atatcttacc tgatgaagga
     1201 gagccaacag atgaagaaac aactggtgac atcagtgatt ccatggattt tgtactgctc
     1261 aactttgcag aaatgaacaa gctctgggtg cgaatgcagc atcagggaca tagccgagat
     1321 agagaaaaaa gagaacgaga aagacaagaa ctgagaattt tagtgggaac aaatttggtg
     1381 cgcctcagtc agttggaagg tgtaaatgtg gaacgttaca aacagattgt tttgactggc
     1441 atattggagc aagttgtaaa ctgtagggat gctttggctc aagaatatct catggagtgt
     1501 attattcagg ttttccctga tgaatttcac ctccagactt tgaatccttt tcttcgggcc
     1561 tgtgctgagt tacaccagaa tgtaaatgtg aagaacataa tcattgcttt aattgataga
     1621 ttagctttat ttgctcaccg tgaagatgga cctggaatcc cagcggatat taaacttttt
     1681 gatatatttt cacagcaggt ggctacagtg atacagtcta gacaagacat gccttcagag
     1741 gatgttgtat ctttacaagt ctctctgatt aatcttgcca tgaaatgtta ccctgatcgt
     1801 gtggactatg ttgataaagt tctagaaaca acagtggaga tattcaataa gctcaacctt
     1861 gaacatattg ctaccagtag tgcagtttca aaggaactca ccagactttt gaaaatacca
     1921 gttgacactt acaacaatat tttaacagtc ttgaaattaa aacattttca cccactcttt
     1981 gagtactttg actacgagtc cagaaagagc atgagttgtt atgtgcttag taatgttctg
     2041 gattataaca cagaaattgt ctctcaagac caggtggatt ccataatgaa tttggtatcc
     2101 acgttgattc aagatcagcc agatcaacct gtagaagacc ctgatccaga agattttgct
     2161 gatgagcaga gccttgtggg ccgcttcatt catctgctgc gctctgagga ccctgaccag
     2221 cagtacttga ttttgaacac agcacgaaaa cattttggag ctggtggaaa tcagcggatt
     2281 cgcttcacac tgccaccttt ggtatttgca gcttaccagc tggcttttcg atataaagag
     2341 aattctaaag tggatgacaa atgggaaaag aaatgccaga agattttttc atttgcccac
     2401 cagactatca gtgctttgat caaagcagag ctggcagaat tgcccttaag actttttctt
     2461 caaggagcac tagctgctgg ggaaattggt tttgaaaatc atgagacagt cgcatatgaa
     2521 ttcatgtccc aggcattttc tctgtatgaa gatgaaatca gcgattccaa agcacagcta
     2581 gctgccatca ccttgatcat tggcactttt gaaaggatga agtgcttcag tgaagagaat
     2641 catgaacctc tgaggactca gtgtgccctt gctgcatcca aacttctaaa gaaacctgat
     2701 cagggccgag ctgtgagcac ctgtgcacat ctcttctggt ctggcagaaa cacggacaaa
     2761 aatggggagg agcttcacgg aggcaagagg gtaatggagt gcctaaaaaa agctctaaaa
     2821 atagcaaatc agtgcatgga cccctctcta caagtgcagc tttttataga aattctgaac
     2881 agatatatct atttttatga aaaggaaaat gatgcggtaa caattcaggt tttaaaccag
     2941 cttatccaaa agattcgaga agacctcccg aatcttgaat ccagtgaaga aacagagcag
     3001 attaacaaac attttcataa cacactggag catttgcgct tgcggcggga atcaccagaa
     3061 tccgaggggc caatttatga aggtctcatc cttggcggat ccatggtgag caagggcgag
     3121 gcagtgatca aggagttcat gcggttcaag gtgcacatgg agggctccat gaacggccac
     3181 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
     3241 ctgaaggtga ccaagggtgg ccccctgccc ttctcctggg acatcctgtc ccctcagttc
     3301 atgtacggct ccagggcctt caccaagcac cccgccgaca tccccgacta ctataagcag
     3361 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgccgtg
     3421 accgtgaccc aggacacctc cctggaggac ggcaccctga tctacaaggt gaagctccgc
     3481 ggcaccaact tccctcctga cggccccgta atgcagaaga agacaatggg ctgggaagcg
     3541 tccaccgagc ggttgtaccc cgaggacggc gtgctgaagg gcgacattaa gatggccctg
     3601 cgcctgaagg acggcggccg ctacctggcg gacttcaaga ccacctacaa ggccaagaag
     3661 cccgtgcaga tgcccggcgc ctacaacgtc gaccgcaagt tggacatcac ctcccacaac
     3721 gaggactaca ccgtggtgga acagtacgaa cgctccgagg gccgccactc caccggcggc
     3781 atggacgagc tgtacaagta gtaactcgag tagacccagc tttcttgtac aaagttggca
     3841 ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata
     3901 aaatcattat ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag
     3961 ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata
     4021 aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
     4081 ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg
     4141 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     4201 gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     4261 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
     4321 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     4381 ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     4441 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     4501 acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
     4561 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     4621 tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     4681 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     4741 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     4801 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     4861 agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact
     4921 ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg
     4981 tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5041 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5101 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5161 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     5221 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5281 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5341 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5401 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5461 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5521 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5581 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5641 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtt
//
