LOCUS       sequence-449417-        8571 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector with Annexin A6 tagged with mScarlet.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8571)
  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 8571)
  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: pEF.DEST51-ANXA6-mScarlet
FEATURES             Location/Qualifiers
     source          1..8571
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(166..185)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        473..1651
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          704..1642
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1599..1619
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1668..1687
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1668..1686
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1718..1742
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3808..4503
                     /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(3941..3959)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4209..4228
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(4519..4543)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4569..4610
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4620..4637
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4660..4677)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4666..4890
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4936..5364
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5023..5042)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5233..5254
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5373..5393)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5378..5707
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5558..5693
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5620..5639
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5755..5802
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5821..6219
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    6377..6498
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6414..6433)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6468..6487
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6547..6563)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6547..6563)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6560..6582)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6571..6587
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6595..6601,6602..6619,6620..6625))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6640..6661
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6778..6795)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6938..7526)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7018..7037)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7697..8488,8489..8557))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8320..8339
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(8558..8571,1..91))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
       61 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
      121 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
      181 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      241 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      301 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      361 ttgattattg actaggcttt tgcaaaaagc tttgcaaaga tggataaagt tttaaacaga
      421 gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa ttggctccgg
      481 tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt
      541 cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt
      601 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc
      661 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg
      721 ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa ttacttccac
      781 ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt gggagagttc
      841 gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct ggcctgggcg
      901 ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa
      961 gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct ggcaagatag
     1021 tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg ccgcgggcgg
     1081 cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc
     1141 accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc ctggcctcgc
     1201 gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac cagttgcgtg
     1261 agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg
     1321 ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc cgtcctcagc
     1381 cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg attagttctc
     1441 gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga tggagtttcc
     1501 ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt aattctcctt
     1561 ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga cagtggttca
     1621 aagttttttt cttccatttc aggtgtcgtg aggaattagc ttggtactaa tacgactcac
     1681 tatagggaga cccaagctgg ctaggtaagc ttgatcaaca agtttgtaca aaaaagcagg
     1741 ctggcgccgg aaccaattca gtcgactggt accggcacca tggccaaacc agcacagggt
     1801 gccaagtacc ggggctccat ccatgacttc ccaggctttg accccaacca ggatgccgag
     1861 gctctgtaca ctgccatgaa gggctttggc agtgacaagg aggccatact ggacataatc
     1921 acctcacgga gcaacaggca gaggcaggag gtctgccaga gctacaagtc cctctacggc
     1981 aaggacctca ttgctgattt aaagtatgaa ttgacgggca agtttgaacg gttgattgtg
     2041 ggcctgatga ggccacctgc ctattgtgat gccaaagaaa ttaaagatgc catctcgggc
     2101 attggcactg atgagaagtg cctcattgag atcttggctt cccggaccaa tgagcagatg
     2161 caccagctgg tggcagcata caaagatgcc tacgagcggg acctggaggc tgacatcatc
     2221 ggcgacacct ctggccactt ccagaagatg cttgtggtcc tgctccaggg aaccagggag
     2281 gaggatgacg tagtgagcga ggacctggta caacaggatg tccaggacct atacgaggca
     2341 ggggaactga aatggggaac agatgaagcc cagttcattt acatcttggg aaatcgcagc
     2401 aagcagcatc ttcggttggt gttcgatgag tatctgaaga ccacagggaa gccgattgaa
     2461 gccagcatcc gaggggagct gtctggggac tttgagaagc taatgctggc cgtagtgaag
     2521 tgtatccgga gcaccccgga atattttgct gaaaggctct tcaaggctat gaagggcctg
     2581 gggactcggg acaacaccct gatccgcatc atggtctccc gtagtgagtt ggacatgctc
     2641 gacattcggg agatcttccg gaccaagtat gagaagtccc tctacagcat gatcaagaat
     2701 gacacctctg gcgagtacaa gaagactctg ctgaagctgt ctgggggaga tgatgatgct
     2761 gctggccagt tcttcccgga ggcagcgcag gtggcctatc agatgtggga acttagtgca
     2821 gtggcccgag tagagctgaa gggaactgtg cgcccagcca atgacttcaa ccctgacgca
     2881 gatgccaaag cgctgcggaa agccatgaag ggactcggga ctgacgaaga cacaatcatc
     2941 gatatcatca cgcaccgcag caatgtccag cggcagcaga tccggcagac cttcaagtct
     3001 cactttggcc gggacttaat gactgacctg aagtctgaga tctctggaga cctggcaagg
     3061 ctgattctgg ggctcatgat gccaccggcc cattacgatg ccaagcagtt gaagaaggcc
     3121 atggagggag ccggcacaga tgaaaaggct cttattgaaa tcctggccac tcggaccaat
     3181 gctgaaatcc gggccatcaa tgaggcctat aaggaggact atcacaagtc cctggaggat
     3241 gctctgagct cagacacatc tggccacttc aggaggatcc tcatttctct ggccacgggg
     3301 catcgtgagg agggaggaga aaacctggac caggcacggg aagatgccca ggtggctgct
     3361 gagatcttgg aaatagcaga cacacctagt ggagacaaaa cttccttgga gacacgtttc
     3421 atgacgatcc tgtgtacccg gagctatccg cacctccgga gagtcttcca ggagttcatc
     3481 aagatgacca actatgacgt ggagcacacc atcaagaagg agatgtctgg ggatgtcagg
     3541 gatgcatttg tggccattgt tcaaagtgtc aagaacaagc ctctcttctt tgccgacaaa
     3601 ctttacaaat ccatgaaggg tgctggcaca gatgagaaga ctctgaccag gatcatggta
     3661 tcccgcagtg agattgacct gctcaacatc cggagggaat tcattgagaa atatgacaag
     3721 tctctccacc aagccattga gggtgacacc tccggagact tcctgaaggc cttgctggct
     3781 ctctgtggtg gtgaggacgg cggatccatg gtgagcaagg gcgaggcagt gatcaaggag
     3841 ttcatgcggt tcaaggtgca catggagggc tccatgaacg gccacgagtt cgagatcgag
     3901 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag
     3961 ggtggccccc tgcccttctc ctgggacatc ctgtcccctc agttcatgta cggctccagg
     4021 gccttcacca agcaccccgc cgacatcccc gactactata agcagtcctt ccccgagggc
     4081 ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg ccgtgaccgt gacccaggac
     4141 acctccctgg aggacggcac cctgatctac aaggtgaagc tccgcggcac caacttccct
     4201 cctgacggcc ccgtaatgca gaagaagaca atgggctggg aagcgtccac cgagcggttg
     4261 taccccgagg acggcgtgct gaagggcgac attaagatgg ccctgcgcct gaaggacggc
     4321 ggccgctacc tggcggactt caagaccacc tacaaggcca agaagcccgt gcagatgccc
     4381 ggcgcctaca acgtcgaccg caagttggac atcacctccc acaacgagga ctacaccgtg
     4441 gtggaacagt acgaacgctc cgagggccgc cactccaccg gcggcatgga cgagctgtac
     4501 aagtagtaac tcgagtagac ccagctttct tgtacaaagt ggttgatcta gagggcccgc
     4561 ggttcgaagg taagcctatc cctaaccctc tcctcggtct cgattctacg cgtaccggtc
     4621 atcatcacca tcaccattga gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     4681 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     4741 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     4801 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     4861 ggcatgctgg ggatgcggtg ggcgctatgg cttctgaggc ggaaagaacc agctggggct
     4921 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     4981 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     5041 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     5101 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     5161 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     5221 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     5281 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     5341 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     5401 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     5461 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     5521 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     5581 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     5641 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     5701 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac
     5761 aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc
     5821 atggccaagc ctttgtctca agaagaatcc accctcattg aaagagcaac ggctacaatc
     5881 aacagcatcc ccatctctga agactacagc gtcgccagcg cagctctctc tagcgacggc
     5941 cgcatcttca ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc agaactcgtg
     6001 gtgctgggca ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga
     6061 aatgagaaca ggggcatctt gagcccctgc ggacggtgcc gacaggtgct tctcgatctg
     6121 catcctggga tcaaagccat agtgaaggac agtgatggac agccgacggc agttgggatt
     6181 cgtgaattgc tgccctctgg ttatgtgtgg gagggctaag cacttcgtgg ccgaggagca
     6241 ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     6301 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     6361 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     6421 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     6481 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     6541 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     6601 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     6661 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     6721 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     6781 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     6841 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     6901 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6961 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     7021 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     7081 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     7141 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     7201 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     7261 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     7321 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     7381 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     7441 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     7501 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     7561 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     7621 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     7681 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     7741 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7801 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7861 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7921 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7981 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     8041 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     8101 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     8161 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     8221 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     8281 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     8341 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     8401 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     8461 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     8521 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt c
//
