LOCUS       sequence-446048-        7370 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway destination vector with N-terminal mScarlet.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7370)
  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 7370)
  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: pDEST-mScarlet-ORF
FEATURES             Location/Qualifiers
     source          1..7370
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             923..1618
                     /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(1056..1074)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1324..1343
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    1640..1764
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(1789..1794,1795..1812,1813..1819)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1873..2532
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1939..1958)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2874..3179
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     3058..3077
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3220..3344)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3378..3395)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3384..3608
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3654..4082
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3741..3760)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3951..3972
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4091..4111)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4096..4425
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4276..4411
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4338..4357
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4473..4520
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4539..4913
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    5043..5164
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5080..5099)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5134..5153
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5213..5229)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5213..5229)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5226..5248)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5237..5253
                     /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(5261..5267,5268..5285,5286..5291))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5306..5327
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5444..5461)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5615..6203)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5695..5714)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6374..7165,7166..7234))
                     /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     6997..7016
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7235..7339)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccatggtgag caagggcgag gcagtgatca aggagttcat
      961 gcggttcaag gtgcacatgg agggctccat gaacggccac gagttcgaga tcgagggcga
     1021 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg
     1081 ccccctgccc ttctcctggg acatcctgtc ccctcagttc atgtacggct ccagggcctt
     1141 caccaagcac cccgccgaca tccccgacta ctataagcag tccttccccg agggcttcaa
     1201 gtgggagcgc gtgatgaact tcgaggacgg cggcgccgtg accgtgaccc aggacacctc
     1261 cctggaggac ggcaccctga tctacaaggt gaagctccgc ggcaccaact tccctcctga
     1321 cggccccgta atgcagaaga agacaatggg ctgggaagcg tccaccgagc ggttgtaccc
     1381 cgaggacggc gtgctgaagg gcgacattaa gatggccctg cgcctgaagg acggcggccg
     1441 ctacctggcg gacttcaaga ccacctacaa ggccaagaag cccgtgcaga tgcccggcgc
     1501 ctacaacgtc gaccgcaagt tggacatcac ctcccacaac gaggactaca ccgtggtgga
     1561 acagtacgaa cgctccgagg gccgccactc caccggcggc atggacgagc tgtacaagat
     1621 cgatggtggc tctggaggta caagtttgta caaaaaagct gaacgagaaa cgtaaaatga
     1681 tataaatatc aatatattaa attagatttt gcataaaaaa cagactacat aatactgtaa
     1741 aacacaacat atccagtcac tatggcggcc gcattaggca ccccaggctt tacactttat
     1801 gcttccggct cgtataatgt gtggattttg agttaggatc cgtcgagatt ttcaggagct
     1861 aaggaagcta aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg
     1921 catcgtaaag aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc
     1981 gttcagctgg atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat
     2041 ccggccttta ttcacattct tgcccgcctg atgaatgctc atccggaatt ccgtatggca
     2101 atgaaagacg gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat
     2161 gagcaaactg aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt
     2221 ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa
     2281 gggtttattg agaatatgtt tttcgtctca gccaatccct gggtgagttt caccagtttt
     2341 gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat
     2401 tatacgcaag gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtttgt
     2461 gatggcttcc atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag
     2521 ggcggggcgt aaagatctgg atccggctta ctaaaagcca gataacagta tgcgtatttg
     2581 cgcgctgatt tttgcggtat aagaatatat actgatatgt atacccgaag tatgtcaaaa
     2641 agaggtatgc tatgaagcag cgtattacag tgacagttga cagcgacagc tatcagttgc
     2701 tcaaggcata tatgatgtca atatctccgg tctggtaagc acaaccatgc agaatgaagc
     2761 ccgtcgtctg cgtgccgaac gctggaaagc ggaaaatcag gaagggatgg ctgaggtcgc
     2821 ccggtttatt gaaatgaacg gctcttttgc tgacgagaac aggggctggt gaaatgcagt
     2881 ttaaggttta cacctataaa agagagagcc gttatcgtct gtttgtggat gtacagagtg
     2941 atattattga cacgcccggg cgacggatgg tgatccccct ggccagtgca cgtctgctgt
     3001 cagataaagt ctcccgtgaa ctttacccgg tggtgcatat cggggatgaa agctggcgca
     3061 tgatgaccac cgatatggcc agtgtgccgg tctccgttat cggggaagaa gtggctgatc
     3121 tcagccaccg cgaaaatgac atcaaaaacg ccattaacct gatgttctgg ggaatataaa
     3181 tgtcaggctc ccttatacac agccagtctg caggtcgacc atagtgactg gatatgttgt
     3241 gttttacagt attatgtagt ctgtttttta tgcaaaatct aatttaatat attgatattt
     3301 atatcatttt acgtttctcg ttcagctttc ttgtacaaag tggttgtcta gagggcccgt
     3361 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     3421 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     3481 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     3541 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     3601 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     3661 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3721 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3781 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3841 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3901 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3961 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     4021 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4081 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4141 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4201 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4261 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4321 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     4381 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     4441 tgtatatcca ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg gcatagtata
     4501 tcggcatagt ataatacgac aaggtgagga actaaaccat ggccaagttg accagtgccg
     4561 ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga gttctggacc gaccggctcg
     4621 ggttctcccg ggacttcgtg gaggacgact tcgccggtgt ggtccgggac gacgtgaccc
     4681 tgttcatcag cgcggtccag gaccaggtgg tgccggacaa caccctggcc tgggtgtggg
     4741 tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg aacttccggg
     4801 acgcctccgg gccggccatg accgagatcg gcgagcagcc gtgggggcgg gagttcgccc
     4861 tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc
     4921 tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4981 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5041 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     5101 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5161 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     5221 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5281 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5341 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5401 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5461 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5521 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5581 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5641 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5701 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5761 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5821 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5881 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5941 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6001 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     6061 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6121 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6181 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6241 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6301 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6361 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6421 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6481 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6541 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6601 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6661 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6721 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6781 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6841 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6901 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6961 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     7021 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7081 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7141 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7201 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7261 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7321 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
