LOCUS       sequence-471960-        5320 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector with Annexin A6 tagged with mScarlet.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5320)
  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 5320)
  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-ANXA6-mScarlet
FEATURES             Location/Qualifiers
     source          1..5320
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     138..157
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(448..547)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(564..583)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(565..583)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(588..604)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(588..604)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(706..725)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             717..1526
                     /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(784..803)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1673..2261
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2162..2181
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2415..2432
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2591..2618)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2710..2796)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2845..2867
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2859..2876
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2860..2876
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2892..2991
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(5057..5320,1..432)
                     /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(5190..5208)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
ORIGIN
        1 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgc cgtgaccgtg
       61 acccaggaca cctccctgga ggacggcacc ctgatctaca aggtgaagct ccgcggcacc
      121 aacttccctc ctgacggccc cgtaatgcag aagaagacaa tgggctggga agcgtccacc
      181 gagcggttgt accccgagga cggcgtgctg aagggcgaca ttaagatggc cctgcgcctg
      241 aaggacggcg gccgctacct ggcggacttc aagaccacct acaaggccaa gaagcccgtg
      301 cagatgcccg gcgcctacaa cgtcgaccgc aagttggaca tcacctccca caacgaggac
      361 tacaccgtgg tggaacagta cgaacgctcc gagggccgcc actccaccgg cggcatggac
      421 gagctgtaca agtagtaact cgagtagacc cagctttctt gtacaaagtt ggcattataa
      481 gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca
      541 ttatttgcca tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt
      601 cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata
      661 tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga
      721 gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat
      781 atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt
      841 atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg
      901 atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca
      961 tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg atccccggaa
     1021 aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc
     1081 tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg
     1141 atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga
     1201 gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata
     1261 aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc
     1321 ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag
     1381 accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac
     1441 agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc
     1501 atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga
     1561 gcattacgct gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt
     1621 acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     1681 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     1741 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     1801 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     1861 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     1921 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     1981 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     2041 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     2101 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     2161 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     2221 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     2281 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     2341 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     2401 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     2461 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     2521 ggaaagcggg cagtgagcgc aacgcaatta atacgcgtac cgctagccag gaagagtttg
     2581 tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttagtttgat gcctggcagt
     2641 ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc
     2701 tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa
     2761 ggcccagtct tccgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg
     2821 cgttaacgct agcatggatg ttttcccagt cacgacgttg taaaacgacg gccagtctta
     2881 agctcgggcc ccaaataatg attttatttt gactgatagt gacctgttcg ttgcaacaca
     2941 ttgatgagca atgctttttt ataatgccaa ctttgtacaa aaaagcaggc tggcgccgga
     3001 accaattcag tcgactggta ccggcaccat ggccaaacca gcacagggtg ccaagtaccg
     3061 gggctccatc catgacttcc caggctttga ccccaaccag gatgccgagg ctctgtacac
     3121 tgccatgaag ggctttggca gtgacaagga ggccatactg gacataatca cctcacggag
     3181 caacaggcag aggcaggagg tctgccagag ctacaagtcc ctctacggca aggacctcat
     3241 tgctgattta aagtatgaat tgacgggcaa gtttgaacgg ttgattgtgg gcctgatgag
     3301 gccacctgcc tattgtgatg ccaaagaaat taaagatgcc atctcgggca ttggcactga
     3361 tgagaagtgc ctcattgaga tcttggcttc ccggaccaat gagcagatgc accagctggt
     3421 ggcagcatac aaagatgcct acgagcggga cctggaggct gacatcatcg gcgacacctc
     3481 tggccacttc cagaagatgc ttgtggtcct gctccaggga accagggagg aggatgacgt
     3541 agtgagcgag gacctggtac aacaggatgt ccaggaccta tacgaggcag gggaactgaa
     3601 atggggaaca gatgaagccc agttcattta catcttggga aatcgcagca agcagcatct
     3661 tcggttggtg ttcgatgagt atctgaagac cacagggaag ccgattgaag ccagcatccg
     3721 aggggagctg tctggggact ttgagaagct aatgctggcc gtagtgaagt gtatccggag
     3781 caccccggaa tattttgctg aaaggctctt caaggctatg aagggcctgg ggactcggga
     3841 caacaccctg atccgcatca tggtctcccg tagtgagttg gacatgctcg acattcggga
     3901 gatcttccgg accaagtatg agaagtccct ctacagcatg atcaagaatg acacctctgg
     3961 cgagtacaag aagactctgc tgaagctgtc tgggggagat gatgatgctg ctggccagtt
     4021 cttcccggag gcagcgcagg tggcctatca gatgtgggaa cttagtgcag tggcccgagt
     4081 agagctgaag ggaactgtgc gcccagccaa tgacttcaac cctgacgcag atgccaaagc
     4141 gctgcggaaa gccatgaagg gactcgggac tgacgaagac acaatcatcg atatcatcac
     4201 gcaccgcagc aatgtccagc ggcagcagat ccggcagacc ttcaagtctc actttggccg
     4261 ggacttaatg actgacctga agtctgagat ctctggagac ctggcaaggc tgattctggg
     4321 gctcatgatg ccaccggccc attacgatgc caagcagttg aagaaggcca tggagggagc
     4381 cggcacagat gaaaaggctc ttattgaaat cctggccact cggaccaatg ctgaaatccg
     4441 ggccatcaat gaggcctata aggaggacta tcacaagtcc ctggaggatg ctctgagctc
     4501 agacacatct ggccacttca ggaggatcct catttctctg gccacggggc atcgtgagga
     4561 gggaggagaa aacctggacc aggcacggga agatgcccag gtggctgctg agatcttgga
     4621 aatagcagac acacctagtg gagacaaaac ttccttggag acacgtttca tgacgatcct
     4681 gtgtacccgg agctatccgc acctccggag agtcttccag gagttcatca agatgaccaa
     4741 ctatgacgtg gagcacacca tcaagaagga gatgtctggg gatgtcaggg atgcatttgt
     4801 ggccattgtt caaagtgtca agaacaagcc tctcttcttt gccgacaaac tttacaaatc
     4861 catgaagggt gctggcacag atgagaagac tctgaccagg atcatggtat cccgcagtga
     4921 gattgacctg ctcaacatcc ggagggaatt cattgagaaa tatgacaagt ctctccacca
     4981 agccattgag ggtgacacct ccggagactt cctgaaggcc ttgctggctc tctgtggtgg
     5041 tgaggacggc ggatccatgg tgagcaaggg cgaggcagtg atcaaggagt tcatgcggtt
     5101 caaggtgcac atggagggct ccatgaacgg ccacgagttc gagatcgagg gcgagggcga
     5161 gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct
     5221 gcccttctcc tgggacatcc tgtcccctca gttcatgtac ggctccaggg ccttcaccaa
     5281 gcaccccgcc gacatccccg actactataa gcagtccttc
//
